关中奶山羊酪蛋白基因启动子表达载体的制作方法

专利名称：关中奶山羊酪蛋白基因启动子表达载体的制作方法
技术领域：
本发明属于生物工程技术领域，涉及一种用克隆关中奶山羊酪蛋白基因启动子区构建的能够使目的蛋白在山羊乳腺中特异性表达并分泌到山羊乳汁中的表达载体。
背景技术：
(一)动物乳腺生物反应器的发展历史与研究现状1980年，Gordon[1]采用显微注射方法将重组DNA导入小鼠受精卵，首次获得了带有外源基因的转基因小鼠，这是人类首次对哺乳动物的遗传信息进行人工改造的尝试。1982年，Palmiter等[2]将大鼠生产激素基因注射到小鼠受精卵中，首次获得了比普通小鼠大得多的“超级小鼠”，并提出可以从转基因动物中提纯有价值药用蛋白的设想。“超级小鼠”诞生这一事实表明，通过性细胞的基因操作可以将外源基因导入哺乳动物的基因组内，并使其得以有效地表达，从而产生相应的生理效应。这一研究结果同时标志着生命科学中一个崭新研究体系一转基因动物体系的形成。
转基因动物被认为是遗传学中继基因连锁分析、体细胞遗传和DNA重组之后的第四代现代生物技术。在“超级小鼠”诞生后的二十多年里，转基因动物研究得到了飞速的发展。目前，转基因小鼠制备技术已经很成熟，并且在生物领域得到了广泛的应用，对生物科学与技术的发展发挥了巨大的推动作用。80年代以后，动物转基因技术逐渐被应用于家兔、家禽、山羊、绵羊、猪、牛等其它动物，目前已有数十种外源基因在上述动物中获得表达，以转基因动物作为生产活性蛋白的工厂(又称动物生物反应器)已经从设想变为现实，并显示出巨大的经济潜力，被誉为二十一世纪的黄金产业。
在动物生物反应器研究中，研究进展最快、开发前景最好的是动物乳腺生物反应器。其基本原理是将具有重要开发价值的目的基因与乳蛋白基因表达调控序列融合，构建成乳腺特异(定点)表达基因构件，将此基因构件导入动物胚胎后获得转基因动物后代。当这些动物产乳时，外源基因在泌乳激素的诱导和乳蛋白基因调控序列的指导下在乳腺中获得表达，表达水平可以接近或超过正常乳蛋白的含量。因此，这种转基因动物的乳腺犹如生产活性蛋白的天然工厂，只要给动物喂以饲料，就能从其乳汁中源源不断地获得具有生物活性的贵重蛋白。
自从重组DNA技术问世以来，人类已经建立了许多基因工程表达系统来生产药用蛋白。虽然这些表达系统都有其各自的优点，但都无法与动物乳腺生物反应器相比。目前使用最广的是原核微生物基因工程表达系统，虽然其工艺流程相对简单、研制周期较短、成本较低，其技术也相当成熟，并已有相当数量的基因工程产品造福人类，但由于原核细胞不能对真核蛋白进行有效的加工、折叠和修饰，复杂的真核基因表达产物往往没有生物活性，或以不溶性的包涵体形式存在，需经过复杂的变性和复性过程才能获得有用产品。目前研究和使用比较多的基因工程另一类系统是动物细胞表达系统，虽然其表达产物的活性较高，但细胞大规模培养的技术和条件要求苛刻，表达水平也不高，价格昂贵，故其实际应用价值受到限制。
转基因动物乳腺生物反应器不仅具有上述表达系统的优点，还可克服它们的缺点和不足，哺乳动物的乳腺表达系统具有以下优点(1)在不损害动物本身健康的情况下，可方便地从乳汁中大量收集目的基因的表达产物。(2)动物具有较高的繁殖能力，可使大规模生产成为可能，且具有生产成本低的优点。(3)乳腺上皮细胞对表达的蛋白具有完善的翻译后加工的能力，如糖基化、磷酸化和羧基化等，从而使表达的蛋白具有与天然蛋白相似的功能。(4)正常哺乳动物乳汁的组成成分已研究得比较清楚，这样便于目的基因表达产物的分离提纯。哺乳动物乳汁中蛋白含量为30～35克/升，一头奶牛每天可以产奶蛋白1000克，一只奶山羊可产奶蛋白200克，而外源基因在动物乳腺中的表达水平可以接近或超过正常乳蛋白的含量[3]。
1990年，荷兰Pharming公司(又称PHP公司)培育成功的表达人乳铁蛋白的转基因牛，每升牛奶中含有人乳铁蛋白1克。乳铁蛋白不仅能够促进婴儿对铁的吸收，而且能够提高婴儿免疫力，抵抗消化道疾病的感染，是母乳的良好替代品。3头转基因奶牛年产牛奶10吨，价值50亿美元。最近，荷兰科学家又成功培育了含有促红细胞生成素(EPO)的转基因牛。1991年，英国爱丁堡PPL制药公司培育成功表达α1-抗胰蛋白酶(AAT)的转基因绵羊[4]。具有抑制弹性蛋白酶的活性，主要用于治疗囊性纤维化(CF)和肺气肿。这种转基因绵羊奶中的AAT含量高达35克/升，从中提纯的基因工程药物已通过II期临床试验，并进入美国市场。目前，该公司已经培育出产生AAT的转基因羊200多只，正在用转基因羊和牛乳腺生物反应器表达的目标产品近20种。美国Genzyme Transgene公司(GTC)与日本的SomitomoMetals合作，共同开发其领先产品凝血酶原III，转基因山羊乳中重组产物的表达量为4g/L，目前已经进入临床试验[3]，部分基因产品见表一。
表一从转基因动物奶液中分离纯化目的蛋白
Clark A J，Bessos H，Bishop J O et al.Bio/Technology，1989，7487～492[14]Wright G，Binicda A，Udell M.J Chem Tech Biotechnol，1994，591I0[15]Harris D P，Andrens A T，Wright G et al.Bioseparation，1997，7(1)31～37[16]Denman J，Hayes M，Oday C dt al.Bio/Technology，1991，9839～843[17]Van Cott K E，Williams B，Velandcr W H et al.J Mol Recognit，1996，9(5～6)407～414[18]Dalton J C，BruLey D F，Kang K A et al.Adv Exp Med Biol，1997，411419～428[19]Degener A，BeLew M，Velander W H et al.J Chromatography，1998，13125～137 Wright G，Carver A，Cottom D et al，Bio/Technology.1991，9830～834[10]Paleyanda R K，Velander W H，Lcc T K et al.Nat Biotechnol，1997，1597I～975英国、美国、荷兰等国家的一些公司利用生物反应器生产抗凝血III、人乳铁蛋白、EPO、干扰素等也已进入临床实验阶段。英国罗斯林研究所利用转基因羊，批量生产治疗肺气肿的血友病的羊奶。我国已获得转人血清白蛋白、胰岛素、干扰素等基因的牛、羊和兔。
中国农业大学教授李宁等采用显微注射法将外源人MAAT(修正的人抗胰蛋白酶)基因导入山羊原核期胚胎内，拟从山羊的奶中，提取人药用蛋白，研发含有人保健蛋白的营养制品和生物药品。有关专家认为，获得人基因转基因羊，标志着我国转基因技术进入了新的阶段，为利用动物乳腺生物反应器生产生物药品探索出了又一个新的途径。国外经济学家预测，大约10年后，转基因动物生产的生物制品就会鼎足于国际市场，单是药物的年销售额就将超过250亿美元。
(二)乳腺特异性表达的基因调控研究动物乳腺特异表达载体的构建及其表达水平的高低在很大程度上取决于对乳蛋白基因表达调控机理的认识。目前，已克隆出许多乳蛋白基因，部分乳蛋白基因及其调控序列的核昔酸己被发表。用于转基因动物乳腺定位表达的调控元件主要有以下四类第一类β-乳球蛋白(BLG)基因调控元件。Simons等将绵羊的BLG基因转入小鼠，绵羊的β-乳球蛋白在小鼠乳腺中特异性表达。其奶液中含量可达23g/L[8]。
第二类酪蛋白基因调控序列。常用牛αS1-酪蛋白基因和羊β-酪蛋白基因的调控序列。如αS1-酪蛋白基因调控序列指导的人白介素-2基因已在转基因兔奶液中成功表达[9]。
第三类乳清酸蛋白(WAP)基因调控序列。WAP是啮齿类动物奶液中的主要蛋白质，在家畜奶液中没有WAP的存在。但WAP基因调控序列可以指导外源基因在家畜奶液中表达[10]。
第四类乳清白蛋白基因调控序列。
尽管不同动物乳汁中乳蛋白的含量不同，但含量最高的都是酪蛋白，在牛奶和羊奶中β-酪蛋白分别约占总乳蛋白的37％和50％以上(Provot et al 1995)，表明β-酪蛋白启动子活性很强，它能启动外源基因在转基因动物的乳腺组织中表达(vi-vito)。因此酪蛋白基因是主要的研究对象。
Gordon K.et al.Genetic transformation of mouse embryos by microinjection of purifiedDNA.PNAS.1980，777380-7348[2]Palmiter RD et al.Dramatic growth of mice that develop from eggs microin-jected withmetallothionein-growth hormone fusiongenes Nature.1982，300611[3]薛京伦 卢大儒 乳腺生物反应器的研究现状，生物技术通报.1998，317-21[4]Wright G，Carver A et al.High level expression of active human alpha-l-antitrypsin inthe milk of transgenic sheep.Bio/Technology.1991，9830-834- --- -[8]Simons J P，Mccienaghan M，Clark A J.Natare，1987，328530～532[9]Buhler T，Bruyere T B，Went D F et al.Bio/Technology，1990，8140～143[10]Paleyanda R K，Velander W H，Lee T K et al.Nat Biotechnol，1997，15971～975(三)、与本发明相关技术上海市儿童医院上海医学遗传研究所曾溢滔教授领导的研究组在遗传学报29(3)；206-211，2002发表关于《山羊β-酪蛋白基因启动子指导的转基因小鼠乳汁高效表达人凝血因子IX》一文中，应用β-酪蛋白基因启动子6.7kb指导外源基因表达，其所构建的山羊β-酪蛋白基因启动子序列在保留原有近端成分外，还保留了上游调控区和β-酪蛋白第一内含子、第一外显子和第二外显子。但将目的蛋白插入载体进行分泌性表达，其效果不理想，原因和结果都需要经过再试验。

发明内容
针对上述现有技术中存在的不足，本发明的目的在于提供一种以西北杨凌特有的高产奶山羊的乳腺组织作为β-酪蛋白基因启动子序列来源的表达载体，此表达载体能够用于建立转基因山羊乳腺生物反应器，使用基因工程手段所构建的目的蛋白特异性地在山羊乳腺中表达、并分泌到山羊乳汁中，以供制备生产目的蛋白。
本发明关中奶山羊酪蛋白基因启动子表达载体，具有商售质粒载体和β-酪蛋白基因启动子及其周围的调控序列以及第一内含子、第一外显子和第二外显子所构成的β-酪蛋白基因启动子区域，其特征在于(1)、所述的β-酪蛋白基因启动子区域序列还包括在第二外显子末端引入作为信号肽的限制性核酸内切酶SgfI位点序列；该启动区域序列为-4359～+2106bp(basepair)，全长6465bp；
(2)、所述的β-酪蛋白基因启动子是以西北杨凌特有的高产关中奶山羊的乳腺组织作为来源，从中提取基因组DNA作为模板，设计合成引物，用高保真的DNA聚合酶进行聚合酶链式反应(PCR)，获得PCR产物，经过拼接、克隆和测定序列而得；(3)、所述的关中奶山羊酪蛋白基因启动子表达载体是将含关中奶山羊β-酪蛋白基因启动区域序列6465bp通过限制性内切酶插入商售的质粒载体中而得。关中奶山羊β-酪蛋白基因启动区域序列全长6465bp如下所示1 AGATGATTTT GCAACCCCCT GCCTCAGGAG ACACTGGGAA ATTTCCTGAG ACATTTTTGATCTACTAAAA CGTTGGGGGA CGGAGTCCTC TGTGACCCTT TAAAGGACTC TGTAAAAACT61 TTCCAAAAGC TGTGCAGTTG GTGCTTCTAC CATCTTCGTG GTAGAGGTCA AGGATGCTGCAAGGTTTTCG ACACGTCAAC CACGAAGATG GTAGAAGCAC CATCTCCAGT TCCTACGACG121 TAAACATTCT ACAACACATT AAGAAAACCC CCACAACAAA GAATTCTTCC GCCAAAAATAATTTGTAAGA TGTTGTGTAA TTCTTTTGGG GGTGTTGTTT CTTAAGAAGG CGGTTTTTAT181 TCAATAATAT GAAGGTTGAA AAATACTGGT CTAGCATGTA GTATGTGCTC AATAGCAAGGAGTTATTATA CTTCCAACTT TTTATGACCA GATCGTACAT CATACACGAG TTATCGTTCC241 AGAGAAAAGA AAGCCTTCCT CACTGATTAA TGCAAAGAAA TAGAGGAAAA CAATAGAATGTCTCTTTTCT TTCGGAAGGA GTGACTAATT ACGTTTCTTT ATCTCCTTTT GTTATCTTAC301 GGAAAGACTA GAGAGCTCTT CAAGCAAATT AGAGATATCA AGGGAACATT TCACGCAAAGCCTTTCTGAT CTCTCGAGAA GTTCGTTTAA TCTCTATAGT TCCCTTGTAA AGTGCGTTTC361 ATGGGCACAA TAAAGGACAG AAATTTTATG GAGGAGTTGC TGATGGAGAG GGAGGCCTGGTACCCGTGTT ATTTCCTGTC TTTAAAATAC CTCCTCAACG ACTACCTCTC CCTCCGGACC421 CGTGCTGCGA TTCCTGGGGT CGCAAAGAGT CGGACACAAC TGAGCGACTG AATTGAACTGGCACGACGCT AAGGACCCCA GCGTTTCTCA GCCTGTGTTG ACTCGCTGAC TTAACTTGAC481 AACTGAACTG GACAAAGCAG AAGATATTAA GAAGAGGTGG TAAGAATACA CAGAAGAACATTGACTTGAC CTGTTTCGTC TTCTATAATT CTTCTCCACC ATTCTTATGT GTCTTCTTGT541 ATATAAAAAA GATCTTCATG ACCCAGATAA CCACGATGAT GTGATCACTC ACCTAGAGCCTATATTTTTT CTAGAAGTAC TGGGTCTATT GGTGCTACTA CACTAGTGAG TGGATCTCGG601 AGACACCCTG GAATGCAAAG TCAAACGGCC TTAGAAAGCC TCACTATGAA CAAAGCTAGTTCTGTGGGAC CTTACGTTTC AGTTTGCCGG AATCTTTCGG AGTGATACTT GTTTCGATCA661 GGAGGTAATG GAATTCCAGT TGAGCTATTT CAAATCTTAA AAGGTGATGC TGTGAAAGTGCCTCCATTAC CTTAAGGTCA ACTCGATAAA GTTTAGAATT TTCCACTACG ACACTTTCAC721 CTGCACTCAA TATGTCAGCA AATTTGGAAA ACTCAGCAGT GGCCACAGGA CTGCCACAATGACGTGAGTT ATACAGTCGT TTAAACCTTT TGAGTCGTCA CCGGTGTCCT GACGGTGTTA781 CCCAAAGAAA AGCAATGACA AAGAATGTTC AAACACCCAC ATGATTGCAC TCATCTCACAGGGTTTCTTT TCGTTACTGT TTCTTACAAG TTTGTGGGTG TACTAACGTG AGTAGAGTGT841 TGCTAGCAAA ATAACTCTCA AAATTCTCCA AGCCAGGCTC CAACAGTACG TGGACCATGAACGATCGTTT TATTGAGAGT TTTAAGAGGT TCGGTCCGAG GTTGTCATGC ACCTGGTACT901 ACTTCCAGAT GTTCAAGCTG GATTTAGAAA AGGCAGAGGA ACCAGAGATC AAATTGCCAATGAAGGTCTA CAAGTTCGAC CTAAATCTTT TCCGTCTCCT TGGTCTCTAG TTTAACGGTT961 CATCCATTGG ATCATCAAAA AAGCACGAGA GTTCCAGAAA AACATCTGCT TTATTGACTAGTAGGTAACC TAGTAGTTTT TTCGTGCTCT CAAGGTCTTT TTGTAGACGA AATAACTGAT1021 CGCTAAAGCC TTTGATTGTG TGGATCACAA TAAACTGTGG AAAATTCTTC AAGAGATGGGGCGATTTCGG AAACTAACAC ACCTAGTGTT ATTTGACACC TTTTAAGAAG TTCTCTACCC1081 AATACCAGAC CACTTTACCT GCCTCCTGAG AAATCTGTAT ACAGGTCCAG AAGCAGCAGTTTATGGTCTG GTGAAATGGA CGGAGGACTC TTTAGACATA TGTCCAGGTC TTCGTCGTCA1141 TAGAACTGGA CATGGAACAA CAGACTGGTT CCAAACTGCG AAAGGGGTAC ATCAAGGAATATCTTGACCT GTACCTTGTT GTCTGACCAA GGTTTGACGC TTTCCCCATG TAGTTCCTTA1201 ATTCATTGGA AGGATTGATG CTGAAGCTGA AACTCCTATA CTTTGGCCAC CTAATGTGAATAAGTAACCT TCCTAACTAC GACTTCGACT TTGAGGATAT GAAACCGGTG GATTACACTT1261 GATCTGACTC ATTGGAAAAG ACTCCAATGC TGGGAAAGAT TGAAGGCAGG AGAAGAGGATCTAGACTGAG TAACCTTTTC TGAGGTTACG ACCCTTTCTA ACTTCCGTCC TCTTCTCCTA1321 GACAGAGGAT GAGATGGTTG GATGGGATCA CTGACTCAAT GGACATGAGT TTGAGTAAGCCTGTCTCCTA CTCTACCAAC CTACCCTAGT GACTGAGTTA CCTGTACTCA AACTCATTCG
1381 TCCAGGGGTT GGTGGTGGAC AGGAAAGCCT GGCGTGCTGC AGTCCACAAG GTCACAAAGAAGGTCCCCAA CCACCACCTG TCCTTTCGGA CCGCACGACG TCAGGTGTTC CAGTGTTTCT1441 TTCGGACATG ACTGAGTGAC TGAACTGATA CTGATGTGCT CAACAAATGT ATCTTGAACTAAGCCTGTAC TGACTCACTG ACTTGACTAT GACTACACGA GTTGTTTACA TAGAACTTGA1501 TGTGTGAAGT TCTATGGTCA CATGTAAAGG AAGAATAATC AGGATTAGCT GTGTGTCTTAACACACTTCA AGATACCAGT GTACATTTCC TTCTTATTAG TCCTAATCGA CACACAGAAT1561 GGAATCAGGG TTCTGAGTTT TATGTGTTCA TAGTATCTGC TGGTTCACAA AACATTTTTCCCTTAGTCCC AAGACTCAAA ATACACAAGT ATCATAGACG ACCAAGTGTT TTGTAAAAAG1621 TTATTCTCTG GTTCTTGATT TACTTTATAA AGTAATCTTA ATAGTTATAC TTCACATAGAAATAAGAGAC CAAGAACTAA ATGAAATATT TCATTAGAAT TATCAATATG AAGTGTATCT1681 TACGAAATTA TTATATTTGG ATAATCTCAT GGAAAGGATT AAATACTCCA TCTATTACGAATGCTTTAAT AATATAAACC TATTAGAGTA CCTTTCCTAA TTTATGAGGT AGATAATGCT1741 GTAATGCTGA ACTATCTACT CCTACCTAAT AATTTGTCAG AATTCACTAA TTCTGTGTTACATTACGACT TGATAGATGA GGATGGATTA TTAAACAGTC TTAAGTGATT AAGACACAAT1801 TATTGTTTCT AAATCTGAAT CATTATATGA ATCCTCAGTA TTTTGTTTTC CTTCCTCTATATAACAAAGA TTTAGACTTA GTAATATACT TAGGAGTCAT AAAACAAAAG GAAGGAGATA1861 ATTTTGGAAT TTATTAAACA GTGCTTCAAA TAATTTTTAG GAAACTGAAG TTTTTAGTAATAAAACCTTA AATAATTTGT CACGAAGTTT ATTAAAAATC CTTTGACTTC AAAAATCATT1921 CAGCTCTATC TCTAAATAGC TTTAGTATCT TGAAAAAGTA ATACAAATTC TCACATCCTTGTCGAGATAG AGATTTATCG AAATCATAGA ACTTTTTCAT TATGTTTAAG AGTGTAGGAA1981 AATTTCCTCT TCTCTAAAAT ATCTTTAAAA TATTCTATGA ATGATATCTC TTAATATTTATTAAAGGAGA AGAGATTTTA TAGAAATTTT ATAAGATACT TACTATAGAG AATTATAAAT2041 TTTTTTTGGC AATCCAACAC AGCTTATGGG ATCTTAGTTC CCCAGTGAGG GATTATATCCAAAAAAACCG TTAGGTTGTG TCGAATACCC TAGAATCAAG GGGTCACTCC CTAATATAGG2101 ATGCCAACTG CAGTGAAAGT ACAAAATCCT AAACTGGACT CACCAGGGAT TTCCCAATATTACGGTTGAC GTCACTTTCA TGTTTTAGGA TTTGACCTGA GTGGTCCCTA AAGGGTTATA2161 CTCCTCTAGT TCTTATTTCT GAATATTTTT GGTCCCTTTA TTGTACTCTT CATCCAACTTGAGGAGATCA AGAATAAAGA CTTATAAAAA CCAGGGAAAT AACATGAGAA GTAGGTTGAA2221 TTCTATTGAT TTCTTTCTTG AGGTTATTAT TTACTTGGTT TCAGTTAGAA ATATATGCAAAAGATAACTA AAGAAAGAAC TCCAATAATA AATGAACCAA AGTCAATCTT TATATACGTT2281 ATCTCAGGAC TGCATATTTC AGATTCATTG GCCAATATGG GAAAAAACCT TTGGCTGAACTAGAGTCCTG ACGTATAAAG TCTAAGTAAC CGGTTATACC CTTTTTTGGA AACCGACTTG2341 AAATCATGCT TATAAAAAAT AGTACTAGAG CATCCTACTT TGACTATATC TTGCTCCTCATTTAGTACGA ATATTTTTTA TCATGATCTC GTAGGATGAA ACTGATATAG AACGAGGAGT2401 TTCAGGGTTA TCTAATACAA TTTCCCCACA TGAAATTCTT TTGCATTATA AAAATGGAAGAAGTCCCAAT AGATTATGTT AAAGGGGTGT ACTTTAAGAA AACGTAATAT TTTTACCTTC2461 CTCTTAGGTA ACATTGCAAA AATTCGAGTT GCTCATATGG CACTTTGCTT CTTACTGGTCGAGAATCCAT TGTAACGTTT TTAAGCTCAA CGAGTATACC GTGAAACGAA GAATGACCAG2521 ATTGTGTTCT GAGGCTTACC TGGACAGGTG GTACCTGATG TCATCTTAAA TTGCTGGCTTTAACACAAGA CTCCGAATGG ACCTGTCCAC CATGGACTAC AGTAGAATTT AACGACCGAA2581 TTTGATTTTC CATTGGACAA GCTTCTTTCT TTAGTATATT GTTAAGGATT TCCTTGATCAAAACTAAAAG GTAACCTGTT CGAAGAAAGA AATCATATAA CAATTCCTAA AGGAACTAGT2641 AGATTTTACC TACTTTTCTG GTCCAATTGG TGAGAGACAG TCATAAGGAA ATGCTGTGTTTCTAAAATGG ATGAAAAGAC CAGGTTAACC ACTCTCTGTC AGTATTCCTT TACGACACAA2701 TATTGCACAA TATGTAAAGC ATCTTCCTGA GAAAATAAAA GGGAAATGTT GAATGGGAAGATAACGTGTT ATACATTTCG TAGAAGGACT CTTTTATTTT CCCTTTACAA CTTACCCTTC2761 GATATGCTTT CTTTTGTATT CCTTTTCTGA GAAATCAAAC TTTTTCACCT GTGGCCTTGGCTATACGAAA GAAAACATAA GGAAAAGACT CTTTAGTTTG AAAAAGTGGA CACCGGAACC2821 CCACCAAAAG CTAACAAATA AAGGCATATG AAGTAGCCAA GGCCTTTTCT AGTTATATCTGGTGGTTTTC GATTGTTTAT TTCCGTATAC TTCATCGGTT CCGGAAAAGA TCAATATAGA2881 ATAACACTGA GTTCATTTCA TCATTTATTT TCCTGACTTC CTCCTGGGTC CATATGAGCATATTGTGACT CAAGTAAAGT AGTAAATAAA AGGACTGAAG GAGGACCCAG GTATACTCGT2941 GTCTTAGAAT GAATATTAGC TGAATAATCC AAATACATAG TAGATGTTGA TTTGGGTTTTCAGAATCTTA CTTATAATCG ACTTATTAGG TTTATGTATC ATCTACAACT AAACCCAAAA3001 CTAAGCAATC CAAGACTTGT ATGACAGTAA GATGTATTAC CATCCAACAC ACATCTCAGCGATTCGTTAG GTTCTGAACA TACTGTCATT CTACATAATG GTAGGTTGTG TGTAGAGTCG3061 ATGATATAAA TGCAAGGTAT ATTGTGAAGA AAAATTTTTA ATTATGTCAA AGTGCTTACTTACTATATTT ACGTTCCATA TAACACTTCT TTTTAAAAAT TAATACAGTT TCACGAATGA
3121 TTAGAAGGTC ATCTATCTGT CCCAAAGCTG TGAATATATA TATTGAAGGT AATGAATAGAAATCTTCCAG TAGATAGACA GGGTTTCGAC ACTTATATAT ATAACTTCCA TTACTTATCT3181 TGAAGCTAAC CTTGTAAAAA TGAGTAGTGT GAAATACAAC TACAATTATG AACATCTGTCACTTCGATTG GAACATTTTT ACTCATCACA CTTTATGTTG ATGTTAATAC TTGTAGACAG3241 ACTAAAGAGG CAAAGAAACT TGAAGATTGC TTTTGCAAAT GGGCTCCTAT TAATAAAAAGTGATTTCTCC GTTTCTTTGA ACTTCTAACG AAAACGTTTA CCCGAGGATA ATTATTTTTC3301 TACTTTTGAG GTCTGGCTCA GACTCTATTG TAGTACTTAG GGTAAGACCC TCCTCCTGTAATGAAAACTC CAGACCGAGT CTGAGATAAC ATCATGAATC CCATTCTGGG AGGAGGACAT3361 TGGGCTTTCA TTTTCTTTCT TGCTTCCCTC ATTTGCCCTT CCATGAATAC TAGCTGATAAACCCGAAAGT AAAAGAAAGA ACGAAGGGAG TAAACGGGAA GGTACTTATG ATCGACTATT3421 ACATTGACTA TAAAAGATAT GAGGCCAAAC TTGAGCTGTC CCATTTTAAT AAATCTGTATTGTAACTGAT ATTTTCTATA CTCCGGTTTG AACTCGACAG GGTAAAATTA TTTAGACATA3481 AAATAATATT TGTTCTACAA AAGTATTATC TAAATAAATG TTACTTTCTG TCTTAAAATCTTTATTATAA ACAAGATGTT TTCATAATAG ATTTATTTAC AATGAAAGAC AGAATTTTAG3541 CCTCAACAAA TCCCCACTAT CTAGAGAATA AGATTGACAT TCCCTGGAAT CACAGCATGCGGAGTTGTTT AGGGGTGATA GATCTCTTAT TCTAACTGTA AGGGACCTTA GTGTCGTACG3601 TTTGTCTGCC ATTATCTGAC CCCTTTCTCT TTCTCTCTTC TCACCTCCAT CTACTCCTTTAAACAGACGG TAATAGACTG GGGAAAGAGA AAGAGAGAAG AGTGGAGGTA GATGAGGAAA3661 TTCCTTGCAA TTCATGACCC AGATTCACTG TTTGATTTGG CTTGCATGTG TGTGTGCTGAAAGGAACGTT AAGTACTGGG TCTAAGTGAC AAACTAAACC GAACGTACAC ACACACGACT3721 GTTGCGTCTG ACTGTTATCA ACCCCATGAA TGATAGTCCA CCAGGCTCTA CTGTCCATGACAACGCAGAC TGACAATAGT TGGGGTACTT ACTATCAGGT GGTCCGAGAT GACAGGTACT3781 AATTTTCCAG TCAAGAATAC TGGAGTGGAT TGCATTTCCT ACTCCATTTG ATTAATTTAGTTAAAAGGTC AGTTCTTATG ACCTCACCTA ACGTAAAGGA TGAGGTAAAC TAATTAAATC3841 TGACTTTTAA ATTTCTTTTT CCATATTCGG GAGCCTATTC TTCCTTTTTA GTCTATACTCACTGAAAATT TAAAGAAAAA GGTATAAGCC CTCGGATAAG AAGGAAAAAT CAGATATGAG3901 TCTTCACTCT TCAGGTCTAA GGTATCATCG TGTGCTTGTT AGCTTGTTAC TTTCTCCATTAGAAGTGAGA AGTCCAGATT CCATAGTAGC ACACGAACAA TCGAACAATG AAAGAGGTAA3961 ATAGCTTAAG CACTAACAAC TGTTCAGGTT GGCATGAAAT TGTGTTCTTT GTGTGGCCTGTATCGAATTC GTGATTGTTG ACAAGTCCAA CCGTACTTTA ACACAAGAAA CACACCGGAC4021 TATATTTCTG TTGTGTATTA GAATTTACCC CAAGATCTCA AAGACCCACT GAATACTAAAATATAAAGAC AACACATAAT CTTAAATGGG GTTCTAGAGT TTCTGGGTGA CTTATGATTT4081 GAGACCTCAT TGTGGTTACA ATAATTTGGG GACTGGGCCA AAACTACCGT GCATCCCAGCCTCTGGAGTA ACACCAATGT TATTAAACCC CTGACCCGGT TTTGATGGCA CGTAGGGTCG4141 CAAGATCTGT AGCTACTGGA CAATTTCATT TCCTTTATCA GATTGTGAGT TATTCCTGTTGTTCTAGACA TCGATGACCT GTTAAAGTAA AGGAAATAGT CTAACACTCA ATAAGGACAA4201 AAAATGCTCC CCAGAATTTC TGGGGACAGA AAAATAGGAA GAATTCATTT CCTAATCATGTTTTACGAGG GGTCTTAAAG ACCCCTGTCT TTTTATCCTT CTTAAGTAAA GGATTAGTAC4261 CAGATTTCTA GGAATTCAAA TCCACTGTTG GTTTTATTTC AAACCACAAA ATTAGCATGCGTCTAAAGAT CCTTAAGTTT AGGTGACAAC CAAAATAAAG TTTGGTGTTT TAATCGTACG4321 CATTAAATAC TATATATAAA CAGCCACTAA ATCAGATCAT TATCCATTCA GCTTCTCCTTGTAATTTATG ATATATATTT GTCGGTGATT TAGTCTAGTA ATAGGTAAGT CGAAGAGGAA4381 CACTTCTTCT CCTCTACTTT GGAAAAAAGG TAAGAATCTC AGATATAATT TCAGGTGTATGTGAAGAAGA GGAGATGAAA CCTTTTTTCC ATTCTTAGAG TCTATATTAA AGTCCACATA4441 CTGCTACTCA TCTTTATTTT GGACTAGGTT AAAATGTAGA AAGAACATAA TTGCTTAAAAGACGATGAGT AGAAATAAAA CCTGATCCAA TTTTACATCT TTCTTGTATT AACGAATTTT4501 TAGATCTTAA AAATAAGGGT GTTTAAGATA AGGTTTACAC TATTTTCAGC AGATATGTTAATCTAGAATT TTTATTCCCA CAAATTCTAT TCCAAATGTG ATAAAAGTCG TCTATACAAT4561 AAAAATAGAA GTGACTATAA AGACTTGATA AAAATTATAG TGACTGCAAA TGTTTTAGGATTTTTATCTT CACTGATATT TCTGAACTAT TTTTAATATC ACTGACGTTT ACAAAATCCT4621 ATATAATAAG ATATAATAAC GGTGGTTGCT ATTTTCTTTA GCACAAGACT AGTTAACAGGTATATTATTC TATATTATTG CCACCAACGA TAAAAGAAAT CGTGTTCTGA TCAATTGTCC4681 CTGTATTAAA AGATCTTTTC TTGAATTAAA TATTTTCAAT TTGATTAAAC CTACCTCAGCGACATAATTT TCTAGAAAAG AACTTAATTT ATAAAAGTTA AACTAATTTG GATGGAGTCG4741 CATAAAGGCA AGCACATTTC ATTTATACTA TGGGGATTTG AATAATTATT ACTGAAGAAGGTATTTCCGT TCGTGTAAAG TAAATATGAT ACCCCTAAAC TTATTAATAA TGACTTCTTC4801 CTCTACCAAC AAAAAGTTTA TAGAGCTATC ATATTTAGTC AAGAGATAAA GAGGGTTGTTGAGATGGTTG TTTTTCAAAT ATCTCGATAG TATAAATCAG TTCTCTATTT CTCCCAACAA
4861 AGGATATATA TGCTATTTGA AAGGTATTTA TAAAAGAAGA GTATATTTAT CAAAATTTCTTCCTATATAT ACGATAAACT TTCCATAAAT ATTTTCTTCT CATATAAATA GTTTTAAAGA4921 CAGAACATCC AAATTTCAAG TTTATCATTT ATCTTACAAT ATTTCAAAAA TATTAAAATAGTCTTGTAGG TTTAAAGTTC AAATAGTAAA TAGAATGTTA TAAAGTTTTT ATAATTTTAT4981 GATACTGAAA TACAGAAGTA AATTAAAGAG AAAGTATTTT ACTTGGTAAA AAAATTCTAGCTATGACTTT ATGTCTTCAT TTAATTTCTC TTTCATAAAA TGAACCATTT TTTTAAGATC5041 GTTGGACAGA GAGTGCCAGG AAACAAAAAC AATGAAAAAT GTGACCTGAC AGGAATTATACAACCTGTCT CTCACGGTCC TTTGTTTTTG TTACTTTTTA CACTGGACTG TCCTTAATAT5101 GCTCAAAGTA TAGTAGTAAG TAATGAAATG GCTTAAAAAT TGGTATATAA AATGCTAGTTCGAGTTTCAT ATCATCATTC ATTACTTTAC CGAATTTTTA ACCATATATT TTACGATCAA5161 ATAAAATAAA CAAAATGCAA TAATATCCTC CCTACATGTA ATGAATTCTA GGTATTATGCTATTTTATTT GTTTTACGTT ATTATAGGAG GGATGTACAT TACTTAAGAT CCATAATACG5221 TCTTTTTGGA AGTCTTGACA ATAAAAATTT TTTTAGAAGT TTATAGGCAT CTTGAATAAAAGAAAAACCT TCAGAACTGT TATTTTTAAA AAAATCTTCA AATATCCGTA GAACTTATTT5281 GTGAAACAAA TTAAGAATTA GTATCCATGA GAAAAATATA GAACAATTTT CCTAATTTAGCACTTTGTTT AATTCTTAAT CATAGGTACT CTTTTTATAT CTTGTTAAAA GGATTAAATC5341 TTTGAAAATC TGGGATTGAA GATGTGTGTC AAGAGATGTT GGTGGCAAGA ACATTTTTTTAAACTTTTAG ACCCTAACTT CTACACACAG TTCTCTACAA CCACCGTTCT TGTAAAAAAA5401 TTCAAGAACT TATAAAAATG CAACAAAACA AACCATTTAA TACATTTTGG TCAAAATCAAAAGTTCTTGA ATATTTTTAC GTTGTTTTGT TTGGTAAATT ATGTAAAACC AGTTTTAGTT5461 TAATGTATTT TATTTTATGC TCCAAGGAGC ATAAAATTGG GGACTGGGCA AGAGAAACTGATTACATAAA ATAAAATACG AGGTTCCTCG TATTTTAACC CCTGACCCGT TCTCTTTGAC5521 ACACCCTGGT AAATTACCAA GAGATAAGTA CACAGTTCTA TGTAGAGAAA ATAAGCATAGTGTGGGACCA TTTAATGGTT CTCTATTCAT GTGTCAAGAT ACATCTCTTT TATTCGTATC5581 TGTATGATCT CTAAAATTAT GTGAGACAAA GGAGAGATGA CATTAGGCAT GTGGGGATGAACATACTAGA GATTTTAATA CACTCTGTTT CCTCTCTACT GTAATCCGTA CACCCCTACT5641 AGACTGAGTA GAGAAGAAAC AATCTAATCA GTCCAAGAAA ACATCTCGAT CAGTGGAACATCTGACTCAT CTCTTCTTTG TTAGATTAGT CAGGTTCTTT TGTAGAGCTA GTCACCTTGT5701 AATAGAAGAA ATGCTAAAAT GAAACAGAAG TCTTACTGGA AATAAAAGAT ATGCATAAGATTATCTTCTT TACGATTTTA CTTTGTCTTC AGAATGACCT TTATTTTCTA TACGTATTCT5761 CAAAAATTCA TGAAAATCAC TTAGTTTAGC AGAGAAAAGA TAAAAATAAA GTATGACCTTGTTTTTAAGT ACTTTTAGTG AATCAAATCG TCTCTTTTCT ATTTTTATTT CATACTGGAA5821 CTTCATATAC ATTGTTTGAT CATATGCACC TCAATAAAAC TGAGTCTCCA ACAGAAATGAGAAGTATATG TAACAAACTA GTATACGTGG AGTTATTTTG ACTCAGAGGT TGTCTTTACT5881 AACATTAATA TTTTGTTCAC TGCTCTAATC CCAGAATCTA AGCGATATCT GGCAATAAAATTGTAATTAT AAAACAAGTG ACGAGATTAG GGTCTTAGAT TCGCTATAGA CCGTTATTTT5941 ATAATAAATA TATATTTTTT AATAAATGAA TCAACCACTT AATTTTTCTG TAAATATCTGTATTATTTAT ATATAAAAAA TTATTTACTT AGTTGGTGAA TTAAAAAGAC ATTTATAGAC6001 TAACTTCTCT TCTGTCTTTC CAAAAACACT CATAAGTACT GTGAATGAGA TGAAAAAGAGATTGAAGAGA AGACAGAAAG GTTTTTGTGA GTATTCATGA CACTTACTCT ACTTTTTCTC6061 TGAAGTAGGA TATAGGCTGT TAGCAGAAAA CATCTGAATG GCTGGCAGTG AAACATTAACACTTCATCCT ATATCCGACA ATCGTCTTTT GTAGACTTAC CGACCGTCAC TTTGTAATTG6121 TTGAAATGTA AGATTAATGA GTAATAGTAA ATTTTAACCT TGGCCATATG ATAAAATGTTAACTTTACAT TCTAATTACT CATTATCATT TAAAATTGGA ACCGGTATAC TATTTTACAA6181 CATTAATATT TTTCTAGAAT ACAGGGCTTT TTGTTTTTGC CATGAGGTTT GCAGGATCTTGTAATTATAA AAAGATCTTA TGTCCCGAAA AACAAAAACG GTACTCCAAA CGTCCTAGAA6241 GGTTCCCTGA CCAGGGATCA AACCTGCACT CCCCTGGAAG CATGGAGTCT TGGACATTTGCCAAGGGACT GGTCCCTAGT TTGGACGTGA GGGGACCTTC GTACCTCAGA ACCTGTAAAC6301 TATTATACAC TATCTTTGGT TCCTTTTAAA GGGAAGTAAT TTTACTTAAA TAAGAAAATAATAATATGTG ATAGAAACCA AGGAAAATTT CCCTTCATTA AAATGAATTT ATTCTTTTAT6361 GATTGACAAG TAATACGCTG TTTCCTCATC TTCCCATTCA CAGGAATCGA GAGCCATGAACTAACTGTTC ATTATGCGAC AAAGGAGTAG AAGGGTAAGT GTCCTTAGCT CTCGGTACTT6421 GGTCCTCATC CTTGCCTGTC TGGTGGCTCT GGCCATTGCG ATCGCCCAGGAGTAG GAACGGACAG ACCACCGAGA CCGGTAACGC TAGCG本发明关中奶山羊β-酪蛋白启动子经过实验检测，证明具有启动子和增强子活性。检测实验及结果如下
1、采取启动子和增强子捕获(promoter and enhancer trap)技术进行检测方法是使用Promega公司商售的荧光素酶报告基因载体pGL3-Basic Vector(其环状结构图见图3)，将本发明6465bp的启动子序列插入其多克隆限制性内切酶位点KpnI和BglII之间，构建成含6465bp启动子序列的质粒pGL3-B65，瞬时转染原代培养的山羊乳腺上皮细胞，在催乳素(prolactin)诱导下，报告基因—荧光素酶的合成量显著增高。实验数据如下

实验结果充分说明所克隆的6465bp DNA序列具有启动子和增强子活性。
2、采取启动子捕获(promoter trap)技术进行检测方法是使用Promega公司商售的荧光素酶报告基因载体pGL3-Enhancer Vector(其环状结构图见附图4)，将本发明6465bp的启动子序列插入其多克隆限制性内切酶位点KpnI和BglII之间，构建成含6465bp的启动子序列的质粒pGL3-E65，瞬时转染原代培养的山羊乳腺上皮细胞，在催乳素(prolactin)诱导下，报告基因—荧光素酶的合成量显著增高。实验数据如下

上述2组数据统计学显著性测验P＜0.05。实验结果表明所克隆的6465bp DNA序列具有启动子活性。


图1是β-酪蛋白启动子区DNA序列示意2是Clontech公司的商售质粒pcDNA3.1(+/-)结构示意3是Promega公司的商售质粒pGL3-Basic Vector结构示意4是Promega公司的商售质粒pGL3-Enhancer Vector结构示意图其中图1本发明所克隆构建的关中奶山羊乳腺β-酪蛋白基因启动区序列包含β-酪蛋白基因上游的启动子区域，编码乳腺β-酪蛋白基因的第一、第二外显子和第一内含子，以及在第二外显子末端引入的限制性核酸内切酶Sgf I位点序列，以利于外源目的基因的插入和表达。
图2中Pcmv为CMV启动子；BGH PA为牛生长激素基因的多聚腺苷酸序列；flori为DNA单链复制起点；SV40ori为SV40病毒复制起点；Neomycin为新霉素抗性基因；Ampicillin为氨苄青霉素抗性基因；SV40pA为SV40病毒多聚腺苷酸序列；PUCori为pUC质粒复制起点；T7 Nhel……Pmel为多限制性内切酶位点序列，是外源DNA插入的位置。本发明将所克隆的β-酪蛋白基因启动区序列6465bp插入pcDNA3.1(-)中的外源DNA插入位置T7 Nhel……Pmel多限制性内切酶位点，即得到本发明所需要的一种关中奶山羊酪蛋白基因启动子表达载体，至限制性内切酶位点XhoI和BamHI之间，构建成真核表达载体。
图3中Ampr为苄青霉素抗性基因；flori为DNA单链复制起点；Synthetic poly(A)signal/transcriptional pause site为多聚腺苷酸合成信号/转录终止点，此序列可降低本底；KpnI.....HindIII为多限制性内切酶位点序列，在此插入待检测的DNA序列；Luc+为荧光素酶基因，是一种报告基因；SV40 late poly(A)signal为SV40病毒晚期转录单元的多聚腺苷酸合成信号；SalI，BamHI，HpaI，XbaI，NarI，NcoI分别是不同的限制性核酸内切酶的位点，并表明其位点的序列位置。
图4中SV40 Enhancer为SV40增强子序列；Ampr为苄青霉素抗性基因；flori为DNA单链复制起点；Synthetic poly(A)signal/transcriptional pause site为多聚腺苷酸合成信号/转录终止点，此序列可降低本底；KpnI.....HindIII为多限制性内切酶位点序列，在此插入待检测的DNA序列；Luc+为荧光素酶基因，是一种报告基因；SV40 latepoly(A)signal为SV40病毒晚期转录单元的多聚腺苷酸合成信号；SalI，BamHI，HpaI，XbaI，NarI，NcoI分别是不同的限制性核酸内切酶的位点，并表明其位点的序列位置。
具体实施例方式
实施例1以Clontech公司的商售质粒pcDNA3.1(-)为例，结合图2，对本发明关中奶山羊酪蛋白基因启动子表达载体作进一步描述从关中奶山羊酪蛋白基因中提取基因组DNA作为模板，设计合成引物，用序列高保的DNA聚合酶进行聚合酶链式反应(简称PCR)扩增，获得的PCR产物，再经过拼接、克隆和测定序列而得到的关中奶山羊β-酪蛋白基因-4359-+2106bp启动区域序列(全长6465bp)；将全长6465bp的序列插入Clontech公司的商售质粒pcDNA3.1(-)中的外源DNA限制性内切酶位点XhoI和BamHI之间，得到本发明所需要的一种关中奶山羊酪蛋白基因启动子表达载体；Clontech公司的商售质粒pcDNA3.1(-)质粒含有Pcmv CMV启动子、BGHPA牛生长激素基因的多聚腺苷酸序列、Neomycin新霉素抗性基因、Ampicillin氨苄青霉素抗性基因、SV40pA病毒多聚腺苷酸序列，其DNA序列全长为5427bp，与所插入的关中奶山羊β-酪蛋白基因启动区域序列6465bp共同构成全长为11838bp的序列，经实际测试，结果与设计完全一致1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTGCACTCT CAGTACAATC TGCTCTGATGCTGCCTAGCC CTCTAGAGGG CTAGGGGATA CCACGTGAGA GTCATGTTAG ACGAGACTAC61 CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCGGGCGTATCAA TTCGGTCATA GACGAGGGAC GAACACACAA CCTCCAGCGA CTCATCACGC121 CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGCGCTCGTTTTA AATTCGATGT TGTTCCGTTC CGAACTGGCT GTTAACGTAC TTCTTAGACG181 TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATTAATCCCAATC CGCAAAACGC GACGAAGCGC TACATGCCCG GTCTATATGC GCAACTGTAA241 GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATACTAATAACTG ATCAATAATT ATCATTAGTT AATGCCCCAG TAATCAAGTA TCGGGTATAT301 TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACCACCTCAAGGC GCAATGTATT GAATGCCATT TACCGGGCGG ACCGACTGGC GGGTTGCTGG361 CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCCGGGCGGGTAA CTGCAGTTAT TACTGCATAC AAGGGTATCA TTGCGGTTAT CCCTGAAAGG421 ATTGACGTCA ATGGGTGGAG TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGTTAACTGCAGT TACCCACCTC ATAAATGCCA TTTGACGGGT GAACCGTCAT GTAGTTCACA481 ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT
TAGTATACGG TTCATGCGGG GGATAACTGC AGTTACTGCC ATTTACCGGG CGGACCGTAA541 ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCATACGGGTCAT GTACTGGAAT ACCCTGAAAG GATGAACCGT CATGTAGATG CATAATCAGT601 TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTGAGCGATAATG GTACCACTAC GCCAAAACCG TCATGTAGTT ACCCGCACCT ATCGCCAAAC661 ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACCTGAGTGCCCC TAAAGGTTCA GAGGTGGGGT AACTGCAGTT ACCCTCAAAC AAAACCGTGG721 AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCGTTTTAGTTGC CCTGAAAGGT TTTACAGCAT TGTTGAGGCG GGGTAACTGC GTTTACCCGC781 GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCACATCCGCACA TGCCACCCTC CAGATATATT CGTCTCGAGA GACCGATTGA TCTCTTGGGT841 CTGCTTACTG GCTTATCGAA ATTAATACGA CTCACTATAG GGAGACCCAA GCTGGCTAGCGACGAATGAC CGAATAGCTT TAATTATGCT GAGTGATATC CCTCTGGGTT CGACCGATCG901 GTTTAAACGG GCCCTCTAGA CAGATGATTT TGCAACCCCC TGCCTCAGGA GACACTGGGACAAATTTGCC CGGGAGATCT GTCTACTAAA ACGTTGGGGG ACGGAGTCCT CTGTGACCCT961 AATTTCCTGA GACATTTTTG ATTCCAAAAG CTGTGCAGTT GGTGCTTCTA CCATCTTCGTTTAAAGGACT CTGTAAAAAC TAAGGTTTTC GACACGTCAA CCACGAAGAT GGTAGAAGCA1021 GGTAGAGGTC AAGGATGCTG CTAAACATTC TACAACACAT TAAGAAAACC CCCACAACAACCATCTCCAG TTCCTACGAC GATTTGTAAG ATGTTGTGTA ATTCTTTTGG GGGTGTTGTT1081 AGAATTCTTC CGCCAAAAAT ATCAATAATA TGAAGGTTGA AAAATACTGG TCTAGCATGTTCTTAAGAAG GCGGTTTTTA TAGTTATTAT ACTTCCAACT TTTTATGACC AGATCGTACA1141 AGTATGTGCT CAATAGCAAG GAGAGAAAAG AAAGCCTTCC TCACTGATTA ATGCAAAGAATCATACACGA GTTATCGTTC CTCTCTTTTC TTTCGGAAGG AGTGACTAAT TACGTTTCTT1201 ATAGAGGAAA ACAATAGAAT GGGAAAGACT AGAGAGCTCT TCAAGCAAAT TAGAGATATCTATCTCCTTT TGTTATCTTA CCCTTTCTGA TCTCTCGAGA AGTTCGTTTA ATCTCTATAG1261 AAGGGAACAT TTCACGCAAA GATGGGCACA ATAAAGGACA GAAATTTTAT GGAGGAGTTGTTCCCTTGTA AAGTGCGTTT CTACCCGTGT TATTTCCTGT CTTTAAAATA CCTCCTCAAC1321 CTGATGGAGA GGGAGGCCTG GCGTGCTGCG ATTCCTGGGG TCGCAAAGAG TCGGACACAAGACTACCTCT CCCTCCGGAC CGCACGACGC TAAGGACCCC AGCGTTTCTC AGCCTGTGTT1381 CTGAGCGACT GAATTGAACT GAACTGAACT GGACAAAGCA GAAGATATTA AGAAGAGGTGGACTCGCTGA CTTAACTTGA CTTGACTTGA CCTGTTTCGT CTTCTATAAT TCTTCTCCAC1441 GTAAGAATAC ACAGAAGAAC AATATAAAAA AGATCTTCAT GACCCAGATA ACCACGATGACATTCTTATG TGTCTTCTTG TTATATTTTT TCTAGAAGTA CTGGGTCTAT TGGTGCTACT1501 TGTGATCACT CACCTAGAGC CAGACACCCT GGAATGCAAA GTCAAACGGC CTTAGAAAGCACACTAGTGA GTGGATCTCG GTCTGTGGGA CCTTACGTTT CAGTTTGCCG GAATCTTTCG1561 CTCACTATGA ACAAAGCTAG TGGAGGTAAT GGAATTCCAG TTGAGCTATT TCAAATCTTAGAGTGATACT TGTTTCGATC ACCTCCATTA CCTTAAGGTC AACTCGATAA AGTTTAGAAT1621 AAAGGTGATG CTGTGAAAGT GCTGCACTCA ATATGTCAGC AAATTTGGAA AACTCAGCAGTTTCCACTAC GACACTTTCA CGACGTGAGT TATACAGTCG TTTAAACCTT TTGAGTCGTC1681 TGGCCACAGG ACTGCCACAA TCCCAAAGAA AAGCAATGAC AAAGAATGTT CAAACACCCAACCGGTGTCC TGACGGTGTT AGGGTTTCTT TTCGTTACTG TTTCTTACAA GTTTGTGGGT1741 CATGATTGCA CTCATCTCAC ATGCTAGCAA AATAACTCTC AAAATTCTCC AAGCCAGGCTGTACTAACGT GAGTAGAGTG TACGATCGTT TTATTGAGAG TTTTAAGAGG TTCGGTCCGA1801 CCAACAGTAC GTGGACCATG AACTTCCAGA TGTTCAAGCT GGATTTAGAA AAGGCAGAGG
GGTTGTCATG CACCTGGTAC TTGAAGGTCT ACAAGTTCGA CCTAAATCTT TTCCGTCTCC1861 AACCAGAGAT CAAATTGCCA ACATCCATTG GATCATCAAA AAAGCACGAG AGTTCCAGAATTGGTCTCTA GTTTAACGGT TGTAGGTAAC CTAGTAGTTT TTTCGTGCTC TCAAGGTCTT1921 AAACATCTGC TTTATTGACT ACGCTAAAGC CTTTGATTGT GTGGATCACA ATAAACTGTGTTTGTAGACG AAATAACTGA TGCGATTTCG GAAACTAACA CACCTAGTGT TATTTGACAC1981 GAAAATTCTT CAAGAGATGG GAATACCAGA CCACTTTACC TGCCTCCTGA GAAATCTGTACTTTTAAGAA GTTCTCTACC CTTATGGTCT GGTGAAATGG ACGGAGGACT CTTTAGACAT2041 TACAGGTCCA GAAGCAGCAG TTAGAACTGG ACATGGAACA ACAGACTGGT TCCAAACTGCATGTCCAGGT CTTCGTCGTC AATCTTGACC TGTACCTTGT TGTCTGACCA AGGTTTGACG2101 GAAAGGGGTA CATCAAGGAA TATTCATTGG AAGGATTGAT GCTGAAGCTG AAACTCCTATCTTTCCCCAT GTAGTTCCTT ATAAGTAACC TTCCTAACTA CGACTTCGAC TTTGAGGATA2161 ACTTTGGCCA CCTAATGTGA AGATCTGACT CATTGGAAAA GACTCCAATG CTGGGAAAGATGAAACCGGT GGATTACACT TCTAGACTGA GTAACCTTTT CTGAGGTTAC GACCCTTTCT2221 TTGAAGGCAG GAGAAGAGGA TGACAGAGGA TGAGATGGTT GGATGGGATC ACTGACTCAAAACTTCCGTC CTCTTCTCCT ACTGTCTCCT ACTCTACCAA CCTACCCTAG TGACTGAGTT2281 TGGACATGAG TTTGAGTAAG CTCCAGGGGT TGGTGGTGGA CAGGAAAGCC TGGCGTGCTGACCTGTACTC AAACTCATTC GAGGTCCCCA ACCACCACCT GTCCTTTCGG ACCGCACGAC2341 CAGTCCACAA GGTCACAAAG ATTCGGACAT GACTGAGTGA CTGAACTGAT ACTGATGTGCGTCAGGTGTT CCAGTGTTTC TAAGCCTGTA CTGACTCACT GACTTGACTA TGACTACACG2401 TCAACAAATG TATCTTGAAC TTGTGTGAAG TTCTATGGTC ACATGTAAAG GAAGAATAATAGTTGTTTAC ATAGAACTTG AACACACTTC AAGATACCAG TGTACATTTC CTTCTTATTA2461 CAGGATTAGC TGTGTGTCTT AGGAATCAGG GTTCTGAGTT TTATGTGTTC ATAGTATCTGGTCCTAATCG ACACACAGAA TCCTTAGTCC CAAGACTCAA AATACACAAG TATCATAGAC2521 CTGGTTCACA AAACATTTTT CTTATTCTCT GGTTCTTGAT TTACTTTATA AAGTAATCTTGACCAAGTGT TTTGTAAAAA GAATAAGAGA CCAAGAACTA AATGAAATAT TTCATTAGAA2581 AATAGTTATA CTTCACATAG ATACGAAATT ATTATATTTG GATAATCTCA TGGAAAGGATTTATCAATAT GAAGTGTATC TATGCTTTAA TAATATAAAC CTATTAGAGT ACCTTTCCTA2641 TAAATACTCC ATCTATTACG AGTAATGCTG AACTATCTAC TCCTACCTAA TAATTTGTCAATTTATGAGG TAGATAATGC TCATTACGAC TTGATAGATG AGGATGGATT ATTAAACAGT2701 GAATTCACTA ATTCTGTGTT ATATTGTTTC TAAATCTGAA TCATTATATG AATCCTCAGTCTTAAGTGAT TAAGACACAA TATAACAAAG ATTTAGACTT AGTAATATAC TTAGGAGTCA2761 ATTTTGTTTT CCTTCCTCTA TATTTTGGAA TTTATTAAAC AGTGCTTCAA ATAATTTTTATAAAACAAAA GGAAGGAGAT ATAAAACCTT AAATAATTTG TCACGAAGTT TATTAAAAAT2821 GGAAACTGAA GTTTTTAGTA ACAGCTCTAT CTCTAAATAG CTTTAGTATC TTGAAAAAGTCCTTTGACTT CAAAAATCAT TGTCGAGATA GAGATTTATC GAAATCATAG AACTTTTTCA2881 AATACAAATT CTCACATCCT TAATTTCCTC TTCTCTAAAA TATCTTTAAA ATATTCTATGTTATGTTTAA GAGTGTAGGA ATTAAAGGAG AAGAGATTTT ATAGAAATTT TATAAGATAC2941 AATGATATCT CTTAATATTT ATTTTTTTGG CAATCCAACA CAGCTTATGG GATCTTAGTTTTACTATAGA GAATTATAAA TAAAAAAACC GTTAGGTTGT GTCGAATACC CTAGAATCAA3001 CCCCAGTGAG GGATTATATC CATGCCAACT GCAGTGAAAG TACAAAATCC TAAACTGGACGGGGTCACTC CCTAATATAG GTACGGTTGA CGTCACTTTC ATGTTTTAGG ATTTGACCTG3061 TCACCAGGGA TTTCCCAATA TCTCCTCTAG TTCTTATTTC TGAATATTTT TGGTCCCTTTAGTGGTCCCT AAAGGGTTAT AGAGGAGATC AAGAATAAAG ACTTATAAAA ACCAGGGAAA3121 ATTGTACTCT TCATCCAACT TTTCTATTGA TTTCTTTCTT GAGGTTATTA TTTACTTGGT
TAACATGAGA AGTAGGTTGA AAAGATAACT AAAGAAAGAA CTCCAATAAT AAATGAACCA3181 TTCAGTTAGA AATATATGCA AATCTCAGGA CTGCATATTT CAGATTCATT GGCCAATATGAAGTCAATCT TTATATACGT TTAGAGTCCT GACGTATAAA GTCTAAGTAA CCGGTTATAC3241 GGAAAAAACC TTTGGCTGAA CAAATCATGC TTATAAAAAA TAGTACTAGA GCATCCTACTCCTTTTTTGG AAACCGACTT GTTTAGTACG AATATTTTTT ATCATGATCT CGTAGGATGA3301 TTGACTATAT CTTGCTCCTC ATTCAGGGTT ATCTAATACA ATTTCCCCAC ATGAAATTCTAACTGATATA GAACGAGGAG TAAGTCCCAA TAGATTATGT TAAAGGGGTG TACTTTAAGA3361 TTTGCATTAT AAAAATGGAA GCTCTTAGGT AACATTGCAA AAATTCGAGT TGCTCATATGAAACGTAATA TTTTTACCTT CGAGAATCCA TTGTAACGTT TTTAAGCTCA ACGAGTATAC3421 GCACTTTGCT TCTTACTGGT CATTGTGTTC TGAGGCTTAC CTGGACAGGT GGTACCTGATCGTGAAACGA AGAATGACCA GTAACACAAG ACTCCGAATG GACCTGTCCA CCATGGACTA3481 GTCATCTTAA ATTGCTGGCT TTTTGATTTT CCATTGGACA AGCTTCTTTC TTTAGTATATCAGTAGAATT TAACGACCGA AAAACTAAAA GGTAACCTGT TCGAAGAAAG AAATCATATA3541 TGTTAAGGAT TTCCTTGATC AAGATTTTAC CTACTTTTCT GGTCCAATTG GTGAGAGACAACAATTCCTA AAGGAACTAG TTCTAAAATG GATGAAAAGA CCAGGTTAAC CACTCTCTGT3601 GTCATAAGGA AATGCTGTGT TTATTGCACA ATATGTAAAG CATCTTCCTG AGAAAATAAACAGTATTCCT TTACGACACA AATAACGTGT TATACATTTC GTAGAAGGAC TCTTTTATTT3661 AGGGAAATGT TGAATGGGAA GGATATGCTT TCTTTTGTAT TCCTTTTCTG AGAAATCAAATCCCTTTACA ACTTACCCTT CCTATACGAA AGAAAACATA AGGAAAAGAC TCTTTAGTTT3721 CTTTTTCACC TGTGGCCTTG GCCACCAAAA GCTAACAAAT AAAGGCATAT GAAGTAGCCAGAAAAAGTGG ACACCGGAAC CGGTGGTTTT CGATTGTTTA TTTCCGTATA CTTCATCGGT3781 AGGCCTTTTC TAGTTATATC TATAACACTG AGTTCATTTC ATCATTTATT TTCCTGACTTTCCGGAAAAG ATCAATATAG ATATTGTGAC TCAAGTAAAG TAGTAAATAA AAGGACTGAA3841 CCTCCTGGGT CCATATGAGC AGTCTTAGAA TGAATATTAG CTGAATAATC CAAATACATAGGAGGACCCA GGTATACTCG TCAGAATCTT ACTTATAATC GACTTATTAG GTTTATGTAT3901 GTAGATGTTG ATTTGGGTTT TCTAAGCAAT CCAAGACTTG TATGACAGTA AGATGTATTACATCTACAAC TAAACCCAAA AGATTCGTTA GGTTCTGAAC ATACTGTCAT TCTACATAAT3961 CCATCCAACA CACATCTCAG CATGATATAA ATGCAAGGTA TATTGTGAAG AAAAATTTTTGGTAGGTTGT GTGTAGAGTC GTACTATATT TACGTTCCAT ATAACACTTC TTTTTAAAAA4021 AATTATGTCA AAGTGCTTAC TTTAGAAGGT CATCTATCTG TCCCAAAGCT GTGAATATATTTAATACAGT TTCACGAATG AAATCTTCCA GTAGATAGAC AGGGTTTCGA CACTTATATA4081 ATATTGAAGG TAATGAATAG ATGAAGCTAA CCTTGTAAAA ATGAGTAGTG TGAAATACAATATAACTTCC ATTACTTATC TACTTCGATT GGAACATTTT TACTCATCAC ACTTTATGTT4141 CTACAATTAT GAACATCTGT CACTAAAGAG GCAAAGAAAC TTGAAGATTG CTTTTGCAAAGATGTTAATA CTTGTAGACA GTGATTTCTC CGTTTCTTTG AACTTCTAAC GAAAACGTTT4201 TGGGCTCCTA TTAATAAAAA GTACTTTTGA GGTCTGGCTC AGACTCTATT GTAGTACTTAACCCGAGGAT AATTATTTTT CATGAAAACT CCAGACCGAG TCTGAGATAA CATCATGAAT4261 GGGTAAGACC CTCCTCCTGT ATGGGCTTTC ATTTTCTTTC TTGCTTCCCT CATTTGCCCTCCCATTCTGG GAGGAGGACA TACCCGAAAG TAAAAGAAAG AACGAAGGGA GTAAACGGGA4321 TCCATGAATA CTAGCTGATA AACATTGACT ATAAAAGATA TGAGGCCAAA CTTGAGCTGTAGGTACTTAT GATCGACTAT TTGTAACTGA TATTTTCTAT ACTCCGGTTT GAACTCGACA4381 CCCATTTTAA TAAATCTGTA TAAATAATAT TTGTTCTACA AAAGTATTAT CTAAATAAATGGGTAAAATT ATTTAGACAT ATTTATTATA AACAAGATGT TTTCATAATA GATTTATTTA4441 GTTACTTTCT GTCTTAAAAT CCCTCAACAA ATCCCCACTA TCTAGAGAAT AAGATTGACA
CAATGAAAGA CAGAATTTTA GGGAGTTGTT TAGGGGTGAT AGATCTCTTA TTCTAACTGT4501 TTCCCTGGAA TCACAGCATG CTTTGTCTGC CATTATCTGA CCCCTTTCTC TTTCTCTCTTAAGGGACCTT AGTGTCGTAC GAAACAGACG GTAATAGACT GGGGAAAGAG AAAGAGAGAA4561 CTCACCTCCA TCTACTCCTT TTTCCTTGCA ATTCATGACC CAGATTCACT GTTTGATTTGGAGTGGAGGT AGATGAGGAA AAAGGAACGT TAAGTACTGG GTCTAAGTGA CAAACTAAAC4621 GCTTGCATGT GTGTGTGCTG AGTTGCGTCT GACTGTTATC AACCCCATGA ATGATAGTCCCGAACGTACA CACACACGAC TCAACGCAGA CTGACAATAG TTGGGGTACT TACTATCAGG4681 ACCAGGCTCT ACTGTCCATG AAATTTTCCA GTCAAGAATA CTGGAGTGGA TTGCATTTCCTGGTCCGAGA TGACAGGTAC TTTAAAAGGT CAGTTCTTAT GACCTCACCT AACGTAAAGG4741 TACTCCATTT GATTAATTTA GTGACTTTTA AATTTCTTTT TCCATATTCG GGAGCCTATTATGAGGTAAA CTAATTAAAT CACTGAAAAT TTAAAGAAAA AGGTATAAGC CCTCGGATAA4801 CTTCCTTTTT AGTCTATACT CTCTTCACTC TTCAGGTCTA AGGTATCATC GTGTGCTTGTGAAGGAAAAA TCAGATATGA GAGAAGTGAG AAGTCCAGAT TCCATAGTAG CACACGAACA4861 TAGCTTGTTA CTTTCTCCAT TATAGCTTAA GCACTAACAA CTGTTCAGGT TGGCATGAAAATCGAACAAT GAAAGAGGTA ATATCGAATT CGTGATTGTT GACAAGTCCA ACCGTACTTT4921 TTGTGTTCTT TGTGTGGCCT GTATATTTCT GTTGTGTATT AGAATTTACC CCAAGATCTCAACACAAGAA ACACACCGGA CATATAAAGA CAACACATAA TCTTAAATGG GGTTCTAGAG4981 AAAGACCCAC TGAATACTAA AGAGACCTCA TTGTGGTTAC AATAATTTGG GGACTGGGCCTTTCTGGGTG ACTTATGATT TCTCTGGAGT AACACCAATG TTATTAAACC CCTGACCCGG5041 AAAACTACCG TGCATCCCAG CCAAGATCTG TAGCTACTGG ACAATTTCAT TTCCTTTATCTTTTGATGGC ACGTAGGGTC GGTTCTAGAC ATCGATGACC TGTTAAAGTA AAGGAAATAG5101 AGATTGTGAG TTATTCCTGT TAAAATGCTC CCCAGAATTT CTGGGGACAG AAAAATAGGATCTAACACTC AATAAGGACA ATTTTACGAG GGGTCTTAAA GACCCCTGTC TTTTTATCCT5161 AGAATTCATT TCCTAATCAT GCAGATTTCT AGGAATTCAA ATCCACTGTT GGTTTTATTTTCTTAAGTAA AGGATTAGTA CGTCTAAAGA TCCTTAAGTT TAGGTGACAA CCAAAATAAA5221 CAAACCACAA AATTAGCATG CCATTAAATA CTATATATAA ACAGCCACTA AATCAGATCAGTTTGGTGTT TTAATCGTAC GGTAATTTAT GATATATATT TGTCGGTGAT TTAGTCTAGT5281 TTATCCATTC AGCTTCTCCT TCACTTCTTC TCCTCTACTT TGGAAAAAAG GTAAGAATCTAATAGGTAAG TCGAAGAGGA AGTGAAGAAG AGGAGATGAA ACCTTTTTTC CATTCTTAGA5341 CAGATATAAT TTCAGGTGTA TCTGCTACTC ATCTTTATTT TGGACTAGGT TAAAATGTAGGTCTATATTA AAGTCCACAT AGACGATGAG TAGAAATAAA ACCTGATCCA ATTTTACATC5401 AAAGAACATA ATTGCTTAAA ATAGATCTTA AAAATAAGGG TGTTTAAGAT AAGGTTTACATTTCTTGTAT TAACGAATTT TATCTAGAAT TTTTATTCCC ACAAATTCTA TTCCAAATGT5461 CTATTTTCAG CAGATATGTT AAAAAATAGA AGTGACTATA AAGACTTGAT AAAAATTATAGATAAAAGTC GTCTATACAA TTTTTTATCT TCACTGATAT TTCTGAACTA TTTTTAATAT5521 GTGACTGCAA ATGTTTTAGG AATATAATAA GATATAATAA CGGTGGTTGC TATTTTCTTTCACTGACGTT TACAAAATCC TTATATTATT CTATATTATT GCCACCAACG ATAAAAGAAA5581 AGCACAAGAC TAGTTAACAG GCTGTATTAA AAGATCTTTT CTTGAATTAA ATATTTTCAATCGTGTTCTG ATCAATTGTC CGACATAATT TTCTAGAAAA GAACTTAATT TATAAAAGTT5641 TTTGATTAAA CCTACCTCAG CCATAAAGGC AAGCACATTT CATTTATACT ATGGGGATTTAAACTAATTT GGATGGAGTC GGTATTTCCG TTCGTGTAAA GTAAATATGA TACCCCTAAA5701 GAATAATTAT TACTGAAGAA GCTCTACCAA CAAAAAGTTT ATAGAGCTAT CATATTTAGTCTTATTAATA ATGACTTCTT CGAGATGGTT GTTTTTCAAA TATCTCGATA GTATAAATCA5761 CAAGAGATAA AGAGGGTTGT TAGGATATAT ATGCTATTTG AAAGGTATTT ATAAAAGAAG
GTTCTCTATT TCTCCCAACA ATCCTATATA TACGATAAAC TTTCCATAAA TATTTTCTTC5821 AGTATATTTA TCAAAATTTC TCAGAACATC CAAATTTCAA GTTTATCATT TATCTTACAATCATATAAAT AGTTTTAAAG AGTCTTGTAG GTTTAAAGTT CAAATAGTAA ATAGAATGTT5881 TATTTCAAAA ATATTAAAAT AGATACTGAA ATACAGAAGT AAATTAAAGA GAAAGTATTTATAAAGTTTT TATAATTTTA TCTATGACTT TATGTCTTCA TTTAATTTCT CTTTCATAAA5941 TACTTGGTAA AAAAATTCTA GGTTGGACAG AGAGTGCCAG GAAACAAAAA CAATGAAAAAATGAACCATT TTTTTAAGAT CCAACCTGTC TCTCACGGTC CTTTGTTTTT GTTACTTTTT6001 TGTGACCTGA CAGGAATTAT AGCTCAAAGT ATAGTAGTAA GTAATGAAAT GGCTTAAAAAACACTGGACT GTCCTTAATA TCGAGTTTCA TATCATCATT CATTACTTTA CCGAATTTTT6061 TTGGTATATA AAATGCTAGT TATAAAATAA ACAAAATGCA ATAATATCCT CCCTACATGTAACCATATAT TTTACGATCA ATATTTTATT TGTTTTACGT TATTATAGGA GGGATGTACA6121 AATGAATTCT AGGTATTATG CTCTTTTTGG AAGTCTTGAC AATAAAAATT TTTTTAGAAGTTACTTAAGA TCCATAATAC GAGAAAAACC TTCAGAACTG TTATTTTTAA AAAAATCTTC6181 TTTATAGGCA TCTTGAATAA AGTGAAACAA ATTAAGAATT AGTATCCATG AGAAAAATATAAATATCCGT AGAACTTATT TCACTTTGTT TAATTCTTAA TCATAGGTAC TCTTTTTATA6241 AGAACAATTT TCCTAATTTA GTTTGAAAAT CTGGGATTGA AGATGTGTGT CAAGAGATGTTCTTGTTAAA AGGATTAAAT CAAACTTTTA GACCCTAACT TCTACACACA GTTCTCTACA6301 TGGTGGCAAG AACATTTTTT TTTCAAGAAC TTATAAAAAT GCAACAAAAC AAACCATTTAACCACCGTTC TTGTAAAAAA AAAGTTCTTG AATATTTTTA CGTTGTTTTG TTTGGTAAAT6361 ATACATTTTG GTCAAAATCA ATAATGTATT TTATTTTATG CTCCAAGGAG CATAAAATTGTATGTAAAAC CAGTTTTAGT TATTACATAA AATAAAATAC GAGGTTCCTC GTATTTTAAC6421 GGGACTGGGC AAGAGAAACT GACACCCTGG TAAATTACCA AGAGATAAGT ACACAGTTCTCCCTGACCCG TTCTCTTTGA CTGTGGGACC ATTTAATGGT TCTCTATTCA TGTGTCAAGA6481 ATGTAGAGAA AATAAGCATA GTGTATGATC TCTAAAATTA TGTGAGACAA AGGAGAGATGTACATCTCTT TTATTCGTAT CACATACTAG AGATTTTAAT ACACTCTGTT TCCTCTCTAC6541 ACATTAGGCA TGTGGGGATG AAGACTGAGT AGAGAAGAAA CAATCTAATC AGTCCAAGAATGTAATCCGT ACACCCCTAC TTCTGACTCA TCTCTTCTTT GTTAGATTAG TCAGGTTCTT6601 AACATCTCGA TCAGTGGAAC AAATAGAAGA AATGCTAAAA TGAAACAGAA GTCTTACTGGTTGTAGAGCT AGTCACCTTG TTTATCTTCT TTACGATTTT ACTTTGTCTT CAGAATGACC6661 AAATAAAAGA TATGCATAAG ACAAAAATTC ATGAAAATCA CTTAGTTTAG CAGAGAAAAGTTTATTTTCT ATACGTATTC TGTTTTTAAG TACTTTTAGT GAATCAAATC GTCTCTTTTC6721 ATAAAAATAA AGTATGACCT TCTTCATATA CATTGTTTGA TCATATGCAC CTCAATAAAATATTTTTATT TCATACTGGA AGAAGTATAT GTAACAAACT AGTATACGTG GAGTTATTTT6781 CTGAGTCTCC AACAGAAATG AAACATTAAT ATTTTGTTCA CTGCTCTAAT CCCAGAATCTGACTCAGAGG TTGTCTTTAC TTTGTAATTA TAAAACAAGT GACGAGATTA GGGTCTTAGA6841 AAGCGATATC TGGCAATAAA AATAATAAAT ATATATTTTT TAATAAATGA ATCAACCACTTTCGCTATAG ACCGTTATTT TTATTATTTA TATATAAAAA ATTATTTACT TAGTTGGTGA6901 TAATTTTTCT GTAAATATCT GTAACTTCTC TTCTGTCTTT CCAAAAACAC TCATAAGTACATTAAAAAGA CATTTATAGA CATTGAAGAG AAGACAGAAA GGTTTTTGTG AGTATTCATG6961 TGTGAATGAG ATGAAAAAGA GTGAAGTAGG ATATAGGCTG TTAGCAGAAA ACATCTGAATACACTTACTC TACTTTTTCT CACTTCATCC TATATCCGAC AATCGTCTTT TGTAGACTTA7021 GGCTGGCAGT GAAACATTAA CTTGAAATGT AAGATTAATG AGTAATAGTA AATTTTAACCCCGACCGTCA CTTTGTAATT GAACTTTACA TTCTAATTAC TCATTATCAT TTAAAATTGG7081 TTGGCCATAT GATAAAATGT TCATTAATAT TTTTCTAGAA TACAGGGCTT TTTGTTTTTG
AACCGGTATA CTATTTTACA AGTAATTATA AAAAGATCTT ATGTCCCGAA AAACAAAAAC7141 CCATGAGGTT TGCAGGATCT TGGTTCCCTG ACCAGGGATC AAACCTGCAC TCCCCTGGAAGGTACTCCAA ACGTCCTAGA ACCAAGGGAC TGGTCCCTAG TTTGGACGTG AGGGGACCTT7201 GCATGGAGTC TTGGACATTT GTATTATACA CTATCTTTGG TTCCTTTTAA AGGGAAGTAACGTACCTCAG AACCTGTAAA CATAATATGT GATAGAAACC AAGGAAAATT TCCCTTCATT7261 TTTTACTTAA ATAAGAAAAT AGATTGACAA GTAATACGCT GTTTCCTCAT CTTCCCATTCAAAATGAATT TATTCTTTTA TCTAACTGTT CATTATGCGA CAAAGGAGTA GAAGGGTAAG7321 ACAGGAATCG AGAGCCATGA AGGTCCTCAT CCTTGCCTGT CTGGTGGCTC TGGCCATTGCTGTCCTTAGC TCTCGGTACT TCCAGGAGTA GGAACGGACA GACCACCGAG ACCGGTAACG7381 GATCGCGGAT CCGAGCTCGG TACCAAGCTT AAGTTTAAAC CCGCTGATCA GCCTCGACTGCTAGCGCCTA GGCTCGAGCC ATGGTTCGAA TTCAAATTTG GGCGACTAGT CGGAGCTGAC7441 TGCCTTCTAG TTGCCAGCCA TCTGTTGTTT GCCCCTCCCC CGTGCCTTCC TTGACCCTGGACGGAAGATC AACGGTCGGT AGACAACAAA CGGGGAGGGG GCACGGAAGG AACTGGGACC7501 AAGGTGCCAC TCCCACTGTC CTTTCCTAAT AAAATGAGGA AATTGCATCG CATTGTCTGATTCCACGGTG AGGGTGACAG GAAAGGATTA TTTTACTCCT TTAACGTAGC GTAACAGACT7561 GTAGGTGTCA TTCTATTCTG GGGGGTGGGG TGGGGCAGGA CAGCAAGGGG GAGGATTGGGCATCCACAGT AAGATAAGAC CCCCCACCCC ACCCCGTCCT GTCGTTCCCC CTCCTAACCC7621 AAGACAATAG CAGGCATGCT GGGGATGCGG TGGGCTCTAT GGCTTCTGAG GCGGAAAGAATTCTGTTATC GTCCGTACGA CCCCTACGCC ACCCGAGATA CCGAAGACTC CGCCTTTCTT7681 CCAGCTGGGG CTCTAGGGGG TATCCCCACG CGCCCTGTAG CGGCGCATTA AGCGCGGCGGGGTCGACCCC GAGATCCCCC ATAGGGGTGC GCGGGACATC GCCGCGTAAT TCGCGCCGCC7741 GTGTGGTGGT TACGCGCAGC GTGACCGCTA CACTTGCCAG CGCCCTAGCG CCCGCTCCTTCACACCACCA ATGCGCGTCG CACTGGCGAT GTGAACGGTC GCGGGATCGC GGGCGAGGAA7801 TCGCTTTCTT CCCTTCCTTT CTCGCCACGT TCGCCGGCTT TCCCCGTCAA GCTCTAAATCAGCGAAAGAA GGGAAGGAAA GAGCGGTGCA AGCGGCCGAA AGGGGCAGTT CGAGATTTAG7861 GGGGGCTCCC TTTAGGGTTC CGATTTAGTG CTTTACGGCA CCTCGACCCC AAAAAACTTGCCCCCGAGGG AAATCCCAAG GCTAAATCAC GAAATGCCGT GGAGCTGGGG TTTTTTGAAC7921 ATTAGGGTGA TGGTTCACGT AGTGGGCCAT CGCCCTGATA GACGGTTTTT CGCCCTTTGATAATCCCACT ACCAAGTGCA TCACCCGGTA GCGGGACTAT CTGCCAAAAA GCGGGAAACT7981 CGTTGGAGTC CACGTTCTTT AATAGTGGAC TCTTGTTCCA AACTGGAACA ACACTCAACCGCAACCTCAG GTGCAAGAAA TTATCACCTG AGAACAAGGT TTGACCTTGT TGTGAGTTGG8041 CTATCTCGGT CTATTCTTTT GATTTATAAG GGATTTTGCC GATTTCGGCC TATTGGTTAAGATAGAGCCA GATAAGAAAA CTAAATATTC CCTAAAACGG CTAAAGCCGG ATAACCAATT8101 AAAATGAGCT GATTTAACAA AAATTTAACG CGAATTAATT CTGTGGAATG TGTGTCAGTTTTTTACTCGA CTAAATTGTT TTTAAATTGC GCTTAATTAA GACACCTTAC ACACAGTCAA8161 AGGGTGTGGA AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAATCCCACACCT TTCAGGGGTC CGAGGGGTCG TCCGTCTTCA TACGTTTCGT ACGTAGAGTT8221 TTAGTCAGCA ACCAGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAGAATCAGTCGT TGGTCCACAC CTTTCAGGGG TCCGAGGGGT CGTCCGTCTT CATACGTTTC8281 CATGCATCTC AATTAGTCAG CAACCATAGT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCTGTACGTAGAG TTAATCAGTC GTTGGTATCA GGGCGGGGAT TGAGGCGGGT AGGGCGGGGA8341 AACTCCGCCC AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGCTTGAGGCGGG TCAAGGCGGG TAAGAGGCGG GGTACCGACT GATTAAAAAA AATAAATACG8401 AGAGGCCGAG GCCGCCTCTG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGG
TCTCCGGCTC CGGCGGAGAC GGAGACTCGA TAAGGTCTTC ATCACTCCTC CGAAAAAACC8461 AGGCCTAGGC TTTTGCAAAA AGCTCCCGGG AGCTTGTATA TCCATTTTCG GATCTGATCATCCGGATCCG AAAACGTTTT TCGAGGGCCC TCGAACATAT AGGTAAAAGC CTAGACTAGT8521 AGAGACAGGA TGAGGATCGT TTCGCATGAT TGAACAAGAT GGATTGCACG CAGGTTCTCCTCTCTGTCCT ACTCCTAGCA AAGCGTACTA ACTTGTTCTA CCTAACGTGC GTCCAAGAGG8581 GGCCGCTTGG GTGGAGAGGC TATTCGGCTA TGACTGGGCA CAACAGACAA TCGGCTGCTCCCGGCGAACC CACCTCTCCG ATAAGCCGAT ACTGACCCGT GTTGTCTGTT AGCCGACGAG8641 TGATGCCGCC GTGTTCCGGC TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG TCAAGACCGAACTACGGCGG CACAAGGCCG ACAGTCGCGT CCCCGCGGGC CAAGAAAAAC AGTTCTGGCT8701 CCTGTCCGGT GCCCTGAATG AACTGCAGGA CGAGGCAGCG CGGCTATCGT GGCTGGCCACGGACAGGCCA CGGGACTTAC TTGACGTCCT GCTCCGTCGC GCCGATAGCA CCGACCGGTG8761 GACGGGCGTT CCTTGCGCAG CTGTGCTCGA CGTTGTCACT GAAGCGGGAA GGGACTGGCTCTGCCCGCAA GGAACGCGTC GACACGAGCT GCAACAGTGA CTTCGCCCTT CCCTGACCGA8821 GCTATTGGGC GAAGTGCCGG GGCAGGATCT CCTGTCATCT CACCTTGCTC CTGCCGAGAACGATAACCCG CTTCACGGCC CCGTCCTAGA GGACAGTAGA GTGGAACGAG GACGGCTCTT8881 AGTATCCATC ATGGCTGATG CAATGCGGCG GCTGCATACG CTTGATCCGG CTACCTGCCCTCATAGGTAG TACCGACTAC GTTACGCCGC CGACGTATGC GAACTAGGCC GATGGACGGG8941 ATTCGACCAC CAAGCGAAAC ATCGCATCGA GCGAGCACGT ACTCGGATGG AAGCCGGTCTTAAGCTGGTG GTTCGCTTTG TAGCGTAGCT CGCTCGTGCA TGAGCCTACC TTCGGCCAGA9001 TGTCGATCAG GATGATCTGG ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG AACTGTTCGCACAGCTAGTC CTACTAGACC TGCTTCTCGT AGTCCCCGAG CGCGGTCGGC TTGACAAGCG9061 CAGGCTCAAG GCGCGCATGC CCGACGGCGA GGATCTCGTC GTGACCCATG GCGATGCCTGGTCCGAGTTC CGCGCGTACG GGCTGCCGCT CCTAGAGCAG CACTGGGTAC CGCTACGGAC9121 CTTGCCGAAT ATCATGGTGG AAAATGGCCG CTTTTCTGGA TTCATCGACT GTGGCCGGCTGAACGGCTTA TAGTACCACC TTTTACCGGC GAAAAGACCT AAGTAGCTGA CACCGGCCGA9181 GGGTGTGGCG GACCGCTATC AGGACATAGC GTTGGCTACC CGTGATATTG CTGAAGAGCTCCCACACCGC CTGGCGATAG TCCTGTATCG CAACCGATGG GCACTATAAC GACTTCTCGA9241 TGGCGGCGAA TGGGCTGACC GCTTCCTCGT GCTTTACGGT ATCGCCGCTC CCGATTCGCAACCGCCGCTT ACCCGACTGG CGAAGGAGCA CGAAATGCCA TAGCGGCGAG GGCTAAGCGT9301 GCGCATCGCC TTCTATCGCC TTCTTGACGA GTTCTTCTGA GCGGGACTCT GGGGTTCGAACGCGTAGCGG AAGATAGCGG AAGAACTGCT CAAGAAGACT CGCCCTGAGA CCCCAAGCTT9361 ATGACCGACC AAGCGACGCC CAACCTGCCA TCACGAGATT TCGATTCCAC CGCCGCCTTCTACTGGCTGG TTCGCTGCGG GTTGGACGGT AGTGCTCTAA AGCTAAGGTG GCGGCGGAAG9421 TATGAAAGGT TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT CCTCCAGCGCATACTTTCCA ACCCGAAGCC TTAGCAAAAG GCCCTGCGGC CGACCTACTA GGAGGTCGCG9481 GGGGATCTCA TGCTGGAGTT CTTCGCCCAC CCCAACTTGT TTATTGCAGC TTATAATGGTCCCCTAGAGT ACGACCTCAA GAAGCGGGTG GGGTTGAACA AATAACGTCG AATATTACCA9541 TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCTATGTTTATTT CGTTATCGTA GTGTTTAAAG TGTTTATTTC GTAAAAAAAG TGACGTAAGA9601 AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATCATG TCTGTATACC GTCGACCTCTTCAACACCAA ACAGGTTTGA GTAGTTACAT AGAATAGTAC AGACATATGG CAGCTGGAGA9661 AGCTAGAGCT TGGCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAATTG TTATCCGCTCTCGATCTCGA ACCGCATTAG TACCAGTATC GACAAAGGAC ACACTTTAAC AATAGGCGAG9721 ACAATTCCAC ACAACATACG AGCCGGAAGC ATAAAGTGTA AAGCCTGGGG TGCCTAATGA
TGTTAAGGTG TGTTGTATGC TCGGCCTTCG TATTTCACAT TTCGGACCCC ACGGATTACT9781 GTGAGCTAAC TCACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTGCACTCGATTG AGTGTAATTA ACGCAACGCG AGTGACGGGC GAAAGGTCAG CCCTTTGGAC9841 TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGGAGCACGGTCG ACGTAATTAC TTAGCCGGTT GCGCGCCCCT CTCCGCCAAA CGCATAACCC9901 CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCGGCGAGAAGGC GAAGGAGCGA GTGACTGAGC GACGCGAGCC AGCAAGCCGA CGCCGCTCGC9961 GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA TAACGCAGGACATAGTCGAG TGAGTTTCCG CCATTATGCC AATAGGTGTC TTAGTCCCCT ATTGCGTCCT10021 AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC CGCGTTGCTGTTCTTGTACA CTCGTTTTCC GGTCGTTTTC CGGTCCTTGG CATTTTTCCG GCGCAACGAC10081 GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG CTCAAGTCAGCGCAAAAAGG TATCCGAGGC GGGGGGACTG CTCGTAGTGT TTTTAGCTGC GAGTTCAGTC10141 AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTCTCCACCGCTT TGGGCTGTCC TGATATTTCT ATGGTCCGCA AAGGGGGACC TTCGAGGGAG10201 GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCGCACGCGAGAG GACAAGGCTG GGACGGCGAA TGGCCTATGG ACAGGCGGAA AGAGGGAAGC10261 GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT GTAGGTCGTTCCTTCGCACC GCGAAAGAGT ATCGAGTGCG ACATCCATAG AGTCAAGCCA CATCCAGCAA10321 CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG CGCCTTATCCGCGAGGTTCG ACCCGACACA CGTGCTTGGG GGGCAAGTCG GGCTGGCGAC GCGGAATAGG10381 GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT GGCAGCAGCCCCATTGATAG CAGAACTCAG GTTGGGCCAT TCTGTGCTGA ATAGCGGTGA CCGTCGTCGG10441 ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGGTGACCATTGT CCTAATCGTC TCGCTCCATA CATCCGCCAC GATGTCTCAA GAACTTCACC10501 TGGCCTAACT ACGGCTACAC TAGAAGAACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCAACCGGATTGA TGCCGATGTG ATCTTCTTGT CATAAACCAT AGACGCGAGA CGACTTCGGT10561 GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC CGCTGGTAGCCAATGGAAGC CTTTTTCTCA ACCATCGAGA ACTAGGCCGT TTGTTTGGTG GCGACCATCG10621 GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCTCCAAAAAAAC AAACGTTCGT CGTCTAATGC GCGTCTTTTT TTCCTAGAGT TCTTCTAGGA10681 TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTGAACTAGAAAA GATGCCCCAG ACTGCGAGTC ACCTTGCTTT TGAGTGCAAT TCCCTAAAAC10741 GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTTCAGTACTCTA ATAGTTTTTC CTAGAAGTGG ATCTAGGAAA ATTTAATTTT TACTTCAAAA10801 AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGTTTTAGTTAGA TTTCATATAT ACTCATTTGA ACCAGACTGT CAATGGTTAC GAATTAGTCA10861 GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTCCTCCGTGGAT AGAGTCGCTA GACAGATAAA GCAAGTAGGT ATCAACGGAC TGAGGGGCAG10921 GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCGCACATCTATT GATGCTATGC CCTCCCGAAT GGTAGACCGG GGTCACGACG TTACTATGGC10981 CGAGACCCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCCGCTCTGGGTG CGAGTGGCCG AGGTCTAAAT AGTCGTTATT TGGTCGGTCG GCCTTCCCGG11041 GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG
CTCGCGTCTT CACCAGGACG TTGAAATAGG CGGAGGTAGG TCAGATAATT AACAACGGCC11101 GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACACTTCGATCTC ATTCATCAAG CGGTCAATTA TCAAACGCGT TGCAACAACG GTAACGATGT11161 GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGACCGTAGCACC ACAGTGCGAG CAGCAAACCA TACCGAAGTA AGTCGAGGCC AAGGGTTGCT11221 TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCTAGTTCCGCTC AATGTACTAG GGGGTACAAC ACGTTTTTTC GCCAATCGAG GAAGCCAGGA11281 CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTGGGCTAGCAAC AGTCTTCATT CAACCGGCGT CACAATAGTG AGTACCAATA CCGTCGTGAC11341 CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCAGTATTAAGAG AATGACAGTA CGGTAGGCAT TCTACGAAAA GACACTGACC ACTCATGAGT11401 ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATATGGTTCAGTA AGACTCTTAT CACATACGCC GCTGGCTCAA CGAGAACGGG CCGCAGTTAT11461 CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCTGCCCTATTAT GGCGCGGTGT ATCGTCTTGA AATTTTCACG AGTAGTAACC TTTTGCAAGA11521 TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACTAGCCCCGCTT TTGAGAGTTC CTAGAATGGC GACAACTCTA GGTCAAGCTA CATTGGGTGA11581 CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAAGCACGTGGGT TGACTAGAAG TCGTAGAAAA TGAAAGTGGT CGCAAAGACC CACTCGTTTT11641 ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTCTGTCCTTCCG TTTTACGGCG TTTTTTCCCT TATTCCCGCT GTGCCTTTAC AACTTATGAG11701 ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGATATGAGAAGG AAAAAGTTAT AATAACTTCG TAAATAGTCC CAATAACAGA GTACTCGCCT11761 TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGAATGTATAAAC TTACATAAAT CTTTTTATTT GTTTATCCCC AAGGCGCGTG TAAAGGGGCT11821 AAAGTGCCAC CTGACGTCTTTCACGGTG GACTGCAG实施例2以Promega公司的商售pGL3-Basic Vector质粒为例，结合图3，对本发明关中奶山羊酪蛋白基因启动子表达载体作进一步描述实施例3以Promega公司的商售pGL3-Enhancer Vector质粒为例，结合图4，对本发明关中奶山羊酪蛋白基因启动子表达载体作进一步描述从关中奶山羊酪蛋白基因中提取基因组DNA作为模板，设计合成引物，用序列高保真的DNA聚合酶进行聚合酶链式反应(简称PCR)扩增，获得的PCR产物，再经过拼接、克隆和测定序列而得到的关中奶山羊β-酪蛋白基因-4359～+2106bp启动区域序列(全长6465bp)；将全长6465bp的序列插入Promega公司的商售pGL3-EnhancerVector质粒中KpnI和BglII多限制性内切酶位点之间，得到本发明所需要的另一种关中奶山羊酪蛋白基因启动子表达载体；Promega公司的商售pGL3-Basic Vector质粒中含有Ampr苄青霉素抗性基因、Luc+为荧光素酶基因、SV40病毒晚期转录单元的多聚腺苷酸合成信号，SV40病毒增强子基因序列。
权利要求
1.关中奶山羊酪蛋白基因启动子表达载体，具有商售质粒载体和β-酪蛋白基因启动子及其周围的调控序列以及第一内含子、第一外显子和第二外显子所构成的β-酪蛋白基因启动子区域，其特征在于(1)、所述的β-酪蛋白基因启动子区域序列还包括在第二外显子末端引入作为信号肽的限制性核酸内切酶Sgf I位点序列；该启动区域序列为-4359～+2106bp，全长6465bp；(2)、所述的β-酪蛋白基因启动子是以西北杨凌特有的高产关中奶山羊的乳腺组织作为来源，从中提取基因组DNA作为模板，设计合成引物，用高保真的DNA聚合酶进行聚合酶链式反应(PCR)，获得的PCR产物，经过拼接、克隆和测定序列而得；(3)、所述的关中奶山羊酪蛋白基因启动子表达载体是将含关中奶山羊β-酪蛋白基因启动区域序列插入商售的质粒载体中后得到的关中奶山羊β-酪蛋白基因启动子表达载体，其DNA序列全长是关中奶山羊β-酪蛋白基因启动区域序列全长6465bp与商售的质粒载体序列全长之和。
2.根据权利要求1所述的关中奶山羊酪蛋白基因启动子表达载体，其特征在于关中奶山羊β-酪蛋白基因启动区域序列全长6465bp如下所示1 AGATGATTTT GCAACCCCCT GCCTCAGGAG ACACTGGGAA ATTTCCTGAG ACATTTTTGATCTACTAAAA CGTTGGGGGA CGGAGTCCTC TGTGACCCTT TAAAGGACTC TGTAAAAACT61 TTCCAAAAGC TGTGCAGTTG GTGCTTCTAC CATCTTCGTG GTAGAGGTCA AGGATGCTGCAAGGTTTTCG ACACGTCAAC CACGAAGATG GTAGAAGCAC CATCTCCAGT TCCTACGACG121 TAAACATTCT ACAACACATT AAGAAAACCC CCACAACAAA GAATTCTTCC GCCAAAAATAATTTGTAAGA TGTTGTGTAA TTCTTTTGGG GGTGTTGTTT CTTAAGAAGG CGGTTTTTAT181 TCAATAATAT GAAGGTTGAA AAATACTGGT CTAGCATGTA GTATGTGCTC AATAGCAAGGAGTTATTATA CTTCCAACTT TTTATGACCA GATCGTACAT CATACACGAG TTATCGTTCC241 AGAGAAAAGA AAGCCTTCCT CACTGATTAA TGCAAAGAAA TAGAGGAAAA CAATAGAATGTCTCTTTTCT TTCGGAAGGA GTGACTAATT ACGTTTCTTT ATCTCCTTTT GTTATCTTAC301 GGAAAGACTA GAGAGCTCTT CAAGCAAATT AGAGATATCA AGGGAACATT TCACGCAAAGCCTTTCTGAT CTCTCGAGAA GTTCGTTTAA TCTCTATAGT TCCCTTGTAA AGTGCGTTTC361 ATGGGCACAA TAAAGGACAG AAATTTTATG GAGGAGTTGC TGATGGAGAG GGAGGCCTGGTACCCGTGTT ATTTCCTGTC TTTAAAATAC CTCCTCAACG ACTACCTCTC CCTCCGGACC421 CGTGCTGCGA TTCCTGGGGT CGCAAAGAGT CGGACACAAC TGAGCGACTG AATTGAACTGGCACGACGCT AAGGACCCCA GCGTTTCTCA GCCTGTGTTG ACTCGCTGAC TTAACTTGAC481 AACTGAACTG GACAAAGCAG AAGATATTAA GAAGAGGTGG TAAGAATACA CAGAAGAACATTGACTTGAC CTGTTTCGTC TTCTATAATT CTTCTCCACC ATTCTTATGT GTCTTCTTGT541 ATATAAAAAA GATCTTCATG ACCCAGATAA CCACGATGAT GTGATCACTC ACCTAGAGCCTATATTTTTT CTAGAAGTAC TGGGTCTATT GGTGCTACTA CACTAGTGAG TGGATCTCGG601 AGACACCCTG GAATGCAAAG TCAAACGGCC TTAGAAAGCC TCACTATGAA CAAAGCTAGTTCTGTGGGAC CTTACGTTTC AGTTTGCCGG AATCTTTCGG AGTGATACTT GTTTCGATCA661 GGAGGTAATG GAATTCCAGT TGAGCTATTT CAAATCTTAA AAGGTGATGC TGTGAAAGTGCCTCCATTAC CTTAAGGTCA ACTCGATAAA GTTTAGAATT TTCCACTACG ACACTTTCAC721 CTGCACTCAA TATGTCAGCA AATTTGGAAA ACTCAGCAGT GGCCACAGGA CTGCCACAATGACGTGAGTT ATACAGTCGT TTAAACCTTT TGAGTCGTCA CCGGTGTCCT GACGGTGTTA781 CCCAAAGAAA AGCAATGACA AAGAATGTTC AAACACCCAC ATGATTGCAC TCATCTCACAGGGTTTCTTT TCGTTACTGT TTCTTACAAG TTTGTGGGTG TACTAACGTG AGTAGAGTGT841 TGCTAGCAAA ATAACTCTCA AAATTCTCCA AGCCAGGCTC CAACAGTACG TGGACCATGAACGATCGTTT TATTGAGAGT TTTAAGAGGT TCGGTCCGAG GTTGTCATGC ACCTGGTACT901 ACTTCCAGAT GTTCAAGCTG GATTTAGAAA AGGCAGAGGA ACCAGAGATC AAATTGCCAATGAAGGTCTA CAAGTTCGAC CTAAATCTTT TCCGTCTCCT TGGTCTCTAG TTTAACGGTT961 CATCCATTGG ATCATCAAAA AAGCACGAGA GTTCCAGAAA AACATCTGCT TTATTGACTAGTAGGTAACC TAGTAGTTTT TTCGTGCTCT CAAGGTCTTT TTGTAGACGA AATAACTGAT1021 CGCTAAAGCC TTTGATTGTG TGGATCACAA TAAACTGTGG AAAATTCTTC AAGAGATGGGGCGATTTCGG AAACTAACAC ACCTAGTGTT ATTTGACACC TTTTAAGAAG TTCTCTACCC1081 AATACCAGAC CACTTTACCT GCCTCCTGAG AAATCTGTAT ACAGGTCCAG AAGCAGCAGTTTATGGTCTG GTGAAATGGA CGGAGGACTC TTTAGACATA TGTCCAGGTC TTCGTCGTCA1141 TAGAACTGGA CATGGAACAA CAGACTGGTT CCAAACTGCG AAAGGGGTAC ATCAAGGAATATCTTGACCT GTACCTTGTT GTCTGACCAA GGTTTGACGC TTTCCCCATG TAGTTCCTTA1201 ATTCATTGGA AGGATTGATG CTGAAGCTGA AACTCCTATA CTTTGGCCAC CTAATGTGAATAAGTAACCT TCCTAACTAC GACTTCGACT TTGAGGATAT GAAACCGGTG GATTACACTT1261 GATCTGACTC ATTGGAAAAG ACTCCAATGC TGGGAAAGAT TGAAGGCAGG AGAAGAGGATCTAGACTGAG TAACCTTTTC TGAGGTTACG ACCCTTTCTA ACTTCCGTCC TCTTCTCCTA1321 GACAGAGGAT GAGATGGTTG GATGGGATCA CTGACTCAAT GGACATGAGT TTGAGTAAGCCTGTCTCCTA CTCTACCAAC CTACCCTAGT GACTGAGTTA CCTGTACTCA AACTCATTCG1381 TCCAGGGGTT GGTGGTGGAC AGGAAAGCCT GGCGTGCTGC AGTCCACAAG GTCACAAAGAAGGTCCCCAA CCACCACCTG TCCTTTCGGA CCGCACGACG TCAGGTGTTC CAGTGTTTCT1441 TTCGGACATG ACTGAGTGAC TGAACTGATA CTGATGTGCT CAACAAATGT ATCTTGAACTAAGCCTGTAC TGACTCACTG ACTTGACTAT GACTACACGA GTTGTTTACA TAGAACTTGA1501 TGTGTGAAGT TCTATGGTCA CATGTAAAGG AAGAATAATC AGGATTAGCT GTGTGTCTTAACACACTTCA AGATACCAGT GTACATTTCC TTCTTATTAG TCCTAATCGA CACACAGAAT1561 GGAATCAGGG TTCTGAGTTT TATGTGTTCA TAGTATCTGC TGGTTCACAA AACATTTTTCCCTTAGTCCC AAGACTCAAA ATACACAAGT ATCATAGACG ACCAAGTGTT TTGTAAAAAG1621 TTATTCTCTG GTTCTTGATT TACTTTATAA AGTAATCTTA ATAGTTATAC TTCACATAGAAATAAGAGAC CAAGAACTAA ATGAAATATT TCATTAGAAT TATCAATATG AAGTGTATCT1681 TACGAAATTA TTATATTTGG ATAATCTCAT GGAAAGGATT AAATACTCCA TCTATTACGAATGCTTTAAT AATATAAACC TATTAGAGTA CCTTTCCTAA TTTATGAGGT AGATAATGCT1741 GTAATGCTGA ACTATCTACT CCTACCTAAT AATTTGTCAG AATTCACTAA TTCTGTGTTACATTACGACT TGATAGATGA GGATGGATTA TTAAACAGTC TTAAGTGATT AAGACACAAT1801 TATTGTTTCT AAATCTGAAT CATTATATGA ATCCTCAGTA TTTTGTTTTC CTTCCTCTATATAACAAAGA TTTAGACTTA GTAATATACT TAGGAGTCAT AAAACAAAAG GAAGGAGATA1861 ATTTTGGAAT TTATTAAACA GTGCTTCAAA TAATTTTTAG GAAACTGAAG TTTTTAGTAATAAAACCTTA AATAATTTGT CACGAAGTTT ATTAAAAATC CTTTGACTTC AAAAATCATT1921 CAGCTCTATC TCTAAATAGC TTTAGTATCT TGAAAAAGTA ATACAAATTC TCACATCCTTGTCGAGATAG AGATTTATCG AAATCATAGA ACTTTTTCAT TATGTTTAAG AGTGTAGGAA1981 AATTTCCTCT TCTCTAAAAT ATCTTTAAAA TATTCTATGA ATGATATCTC TTAATATTTATTAAAGGAGA AGAGATTTTA TAGAAATTTT ATAAGATACT TACTATAGAG AATTATAAAT2041 TTTTTTTGGC AATCCAACAC AGCTTATGGG ATCTTAGTTC CCCAGTGAGG GATTATATCCAAAAAAACCG TTAGGTTGTG TCGAATACCC TAGAATCAAG GGGTCACTCC CTAATATAGG2101 ATGCCAACTG CAGTGAAAGT ACAAAATCCT AAACTGGACT CACCAGGGAT TTCCCAATATTACGGTTGAC GTCACTTTCA TGTTTTAGGA TTTGACCTGA GTGGTCCCTA AAGGGTTATA2161 CTCCTCTAGT TCTTATTTCT GAATATTTTT GGTCCCTTTA TTGTACTCTT CATCCAACTTGAGGAGATCA AGAATAAAGA CTTATAAAAA CCAGGGAAAT AACATGAGAA GTAGGTTGAA2221 TTCTATTGAT TTCTTTCTTG AGGTTATTAT TTACTTGGTT TCAGTTAGAA ATATATGCAAAAGATAACTA AAGAAAGAAC TCCAATAATA AATGAACCAA AGTCAATCTT TATATACGTT2281 ATCTCAGGAC TGCATATTTC AGATTCATTG GCCAATATGG GAAAAAACCT TTGGCTGAACTAGAGTCCTG ACGTATAAAG TCTAAGTAAC CGGTTATACC CTTTTTTGGA AACCGACTTG2341 AAATCATGCT TATAAAAAAT AGTACTAGAG CATCCTACTT TGACTATATC TTGCTCCTCATTTAGTACGA ATATTTTTTA TCATGATCTC GTAGGATGAA ACTGATATAG AACGAGGAGT2401 TTCAGGGTTA TCTAATACAA TTTCCCCACA TGAAATTCTT TTGCATTATA AAAATGGAAGAAGTCCCAAT AGATTATGTT AAAGGGGTGT ACTTTAAGAA AACGTAATAT TTTTACCTTC2461 CTCTTAGGTA ACATTGCAAA AATTCGAGTT GCTCATATGG CACTTTGCTT CTTACTGGTCGAGAATCCAT TGTAACGTTT TTAAGCTCAA CGAGTATACC GTGAAACGAA GAATGACCAG2521 ATTGTGTTCT GAGGCTTACC TGGACAGGTG GTACCTGATG TCATCTTAAA TTGCTGGCTTTAACACAAGA CTCCGAATGG ACCTGTCCAC CATGGACTAC AGTAGAATTT AACGACCGAA2581 TTTGATTTTC CATTGGACAA GCTTCTTTCT TTAGTATATT GTTAAGGATT TCCTTGATCAAAACTAAAAG GTAACCTGTT CGAAGAAAGA AATCATATAA CAATTCCTAA AGGAACTAGT2641 AGATTTTACC TACTTTTCTG GTCCAATTGG TGAGAGACAG TCATAAGGAA ATGCTGTGTTTCTAAAATGG ATGAAAAGAC CAGGTTAACC ACTCTCTGTC AGTATTCCTT TACGACACAA2701 TATTGCACAA TATGTAAAGC ATCTTCCTGA GAAAATAAAA GGGAAATGTT GAATGGGAAGATAACGTGTT ATACATTTCG TAGAAGGACT CTTTTATTTT CCCTTTACAA CTTACCCTTC2761 GATATGCTTT CTTTTGTATT CCTTTTCTGA GAAATCAAAC TTTTTCACCT GTGGCCTTGGCTATACGAAA GAAAACATAA GGAAAAGACT CTTTAGTTTG AAAAAGTGGA CACCGGAACC2821 CCACCAAAAG CTAACAAATA AAGGCATATG AAGTAGCCAA GGCCTTTTCT AGTTATATCTGGTGGTTTTC GATTGTTTAT TTCCGTATAC TTCATCGGTT CCGGAAAAGA TCAATATAGA2881 ATAACACTGA GTTCATTTCA TCATTTATTT TCCTGACTTC CTCCTGGGTC CATATGAGCATATTGTGACT CAAGTAAAGT AGTAAATAAA AGGACTGAAG GAGGACCCAG GTATACTCGT2941 GTCTTAGAAT GAATATTAGC TGAATAATCC AAATACATAG TAGATGTTGA TTTGGGTTTTCAGAATCTTA CTTATAATCG ACTTATTAGG TTTATGTATC ATCTACAACT AAACCCAAAA3001 CTAAGCAATC CAAGACTTGT ATGACAGTAA GATGTATTAC CATCCAACAC ACATCTCAGCGATTCGTTAG GTTCTGAACA TACTGTCATT CTACATAATG GTAGGTTGTG TGTAGAGTCG3061 ATGATATAAA TGCAAGGTAT ATTGTGAAGA AAAATTTTTA ATTATGTCAA AGTGCTTACTTACTATATTT ACGTTCCATA TAACACTTCT TTTTAAAAAT TAATACAGTT TCACGAATGA3121 TTAGAAGGTC ATCTATCTGT CCCAAAGCTG TGAATATATA TATTGAAGGT AATGAATAGAAATCTTCCAG TAGATAGACA GGGTTTCGAC ACTTATATAT ATAACTTCCA TTACTTATCT3181 TGAAGCTAAC CTTGTAAAAA TGAGTAGTGT GAAATACAAC TACAATTATG AACATCTGTCACTTCGATTG GAACATTTTT ACTCATCACA CTTTATGTTG ATGTTAATAC TTGTAGACAG3241 ACTAAAGAGG CAAAGAAACT TGAAGATTGC TTTTGCAAAT GGGCTCCTAT TAATAAAAAGTGATTTCTCC GTTTCTTTGA ACTTCTAACG AAAACGTTTA CCCGAGGATA ATTATTTTTC3301 TACTTTTGAG GTCTGGCTCA GACTCTATTG TAGTACTTAG GGTAAGACCC TCCTCCTGTAATGAAAACTC CAGACCGAGT CTGAGATAAC ATCATGAATC CCATTCTGGG AGGAGGACAT3361 TGGGCTTTCA TTTTCTTTCT TGCTTCCCTC ATTTGCCCTT CCATGAATAC TAGCTGATAAACCCGAAAGT AAAAGAAAGA ACGAAGGGAG TAAACGGGAA GGTACTTATG ATCGACTATT3421 ACATTGACTA TAAAAGATAT GAGGCCAAAC TTGAGCTGTC CCATTTTAAT AAATCTGTATTGTAACTGAT ATTTTCTATA CTCCGGTTTG AACTCGACAG GGTAAAATTA TTTAGACATA3481 AAATAATATT TGTTCTACAA AAGTATTATC TAAATAAATG TTACTTTCTG TCTTAAAATCTTTATTATAA ACAAGATGTT TTCATAATAG ATTTATTTAC AATGAAAGAC AGAATTTTAG3541 CCTCAACAAA TCCCCACTAT CTAGAGAATA AGATTGACAT TCCCTGGAAT CACAGCATGCGGAGTTGTTT AGGGGTGATA GATCTCTTAT TCTAACTGTA AGGGACCTTA GTGTCGTACG3601 TTTGTCTGCC ATTATCTGAC CCCTTTCTCT TTCTCTCTTC TCACCTCCAT CTACTCCTTTAAACAGACGG TAATAGACTG GGGAAAGAGA AAGAGAGAAG AGTGGAGGTA GATGAGGAAA3661 TTCCTTGCAA TTCATGACCC AGATTCACTG TTTGATTTGG CTTGCATGTG TGTGTGCTGAAAGGAACGTT AAGTACTGGG TCTAAGTGAC AAACTAAACC GAACGTACAC ACACACGACT3721 GTTGCGTCTG ACTGTTATCA ACCCCATGAA TGATAGTCCA CCAGGCTCTA CTGTCCATGACAACGCAGAC TGACAATAGT TGGGGTACTT ACTATCAGGT GGTCCGAGAT GACAGGTACT3781 AATTTTCCAG TCAAGAATAC TGGAGTGGAT TGCATTTCCT ACTCCATTTG ATTAATTTAGTTAAAAGGTC AGTTCTTATG ACCTCACCTA ACGTAAAGGA TGAGGTAAAC TAATTAAATC3841 TGACTTTTAA ATTTCTTTTT CCATATTCGG GAGCCTATTC TTCCTTTTTA GTCTATACTCACTGAAAATT TAAAGAAAAA GGTATAAGCC CTCGGATAAG AAGGAAAAAT CAGATATGAG3901 TCTTCACTCT TCAGGTCTAA GGTATCATCG TGTGCTTGTT AGCTTGTTAC TTTCTCCATTAGAAGTGAGA AGTCCAGATT CCATAGTAGC ACACGAACAA TCGAACAATG AAAGAGGTAA3961 ATAGCTTAAG CACTAACAAC TGTTCAGGTT GGCATGAAAT TGTGTTCTTT GTGTGGCCTGTATCGAATTC GTGATTGTTG ACAAGTCCAA CCGTACTTTA ACACAAGAAA CACACCGGAC4021 TATATTTCTG TTGTGTATTA GAATTTACCC CAAGATCTCA AAGACCCACT GAATACTAAAATATAAAGAC AACACATAAT CTTAAATGGG GTTCTAGAGT TTCTGGGTGA CTTATGATTT4081 GAGACCTCAT TGTGGTTACA ATAATTTGGG GACTGGGCCA AAACTACCGT GCATCCCAGCCTCTGGAGTA ACACCAATGT TATTAAACCC CTGACCCGGT TTTGATGGCA CGTAGGGTCG4141 CAAGATCTGT AGCTACTGGA CAATTTCATT TCCTTTATCA GATTGTGAGT TATTCCTGTTGTTCTAGACA TCGATGACCT GTTAAAGTAA AGGAAATAGT CTAACACTCA ATAAGGACAA4201 AAAATGCTCC CCAGAATTTC TGGGGACAGA AAAATAGGAA GAATTCATTT CCTAATCATGTTTTACGAGG GGTCTTAAAG ACCCCTGTCT TTTTATCCTT CTTAAGTAAA GGATTAGTAC4261 CAGATTTCTA GGAATTCAAA TCCACTGTTG GTTTTATTTC AAACCACAAA ATTAGCATGCGTCTAAAGAT CCTTAAGTTT AGGTGACAAC CAAAATAAAG TTTGGTGTTT TAATCGTACG4321 CATTAAATAC TATATATAAA CAGCCACTAA ATCAGATCAT TATCCATTCA GCTTCTCCTTGTAATTTATG ATATATATTT GTCGGTGATT TAGTCTAGTA ATAGGTAAGT CGAAGAGGAA4381 CACTTCTTCT CCTCTACTTT GGAAAAAAGG TAAGAATCTC AGATATAATT TCAGGTGTATGTGAAGAAGA GGAGATGAAA CCTTTTTTCC ATTCTTAGAG TCTATATTAA AGTCCACATA4441 CTGCTACTCA TCTTTATTTT GGACTAGGTT AAAATGTAGA AAGAACATAA TTGCTTAAAAGACGATGAGT AGAAATAAAA CCTGATCCAA TTTTACATCT TTCTTGTATT AACGAATTTT4501 TAGATCTTAA AAATAAGGGT GTTTAAGATA AGGTTTACAC TATTTTCAGC AGATATGTTAATCTAGAATT TTTATTCCCA CAAATTCTAT TCCAAATGTG ATAAAAGTCG TCTATACAAT4561 AAAAATAGAA GTGACTATAA AGACTTGATA AAAATTATAG TGACTGCAAA TGTTTTAGGATTTTTATCTT CACTGATATT TCTGAACTAT TTTTAATATC ACTGACGTTT ACAAAATCCT4621 ATATAATAAG ATATAATAAC GGTGGTTGCT ATTTTCTTTA GCACAAGACT AGTTAACAGGTATATTATTC TATATTATTG CCACCAACGA TAAAAGAAAT CGTGTTCTGA TCAATTGTCC4681 CTGTATTAAA AGATCTTTTC TTGAATTAAA TATTTTCAAT TTGATTAAAC CTACCTCAGCGACATAATTT TCTAGAAAAG AACTTAATTT ATAAAAGTTA AACTAATTTG GATGGAGTCG4741 CATAAAGGCA AGCACATTTC ATTTATACTA TGGGGATTTG AATAATTATT ACTGAAGAAGGTATTTCCGT TCGTGTAAAG TAAATATGAT ACCCCTAAAC TTATTAATAA TGACTTCTTC4801 CTCTACCAAC AAAAAGTTTA TAGAGCTATC ATATTTAGTC AAGAGATAAA GAGGGTTGTTGAGATGGTTG TTTTTCAAAT ATCTCGATAG TATAAATCAG TTCTCTATTT CTCCCAACAA4861 AGGATATATA TGCTATTTGA AAGGTATTTA TAAAAGAAGA GTATATTTAT CAAAATTTCTTCCTATATAT ACGATAAACT TTCCATAAAT ATTTTCTTCT CATATAAATA GTTTTAAAGA4921 CAGAACATCC AAATTTCAAG TTTATCATTT ATCTTACAAT ATTTCAAAAA TATTAAAATAGTCTTGTAGG TTTAAAGTTC AAATAGTAAA TAGAATGTTA TAAAGTTTTT ATAATTTTAT4981 GATACTGAAA TACAGAAGTA AATTAAAGAG AAAGTATTTT ACTTGGTAAA AAAATTCTAGCTATGACTTT ATGTCTTCAT TTAATTTCTC TTTCATAAAA TGAACCATTT TTTTAAGATC5041 GTTGGACAGA GAGTGCCAGG AAACAAAAAC AATGAAAAAT GTGACCTGAC AGGAATTATACAACCTGTCT CTCACGGTCC TTTGTTTTTG TTACTTTTTA CACTGGACTG TCCTTAATAT5101 GCTCAAAGTA TAGTAGTAAG TAATGAAATG GCTTAAAAAT TGGTATATAA AATGCTAGTTCGAGTTTCAT ATCATCATTC ATTACTTTAC CGAATTTTTA ACCATATATT TTACGATCAA5161 ATAAAATAAA CAAAATGCAA TAATATCCTC CCTACATGTA ATGAATTCTA GGTATTATGCTATTTTATTT GTTTTACGTT ATTATAGGAG GGATGTACAT TACTTAAGAT CCATAATACG5221 TCTTTTTGGA AGTCTTGACA ATAAAAATTT TTTTAGAAGT TTATAGGCAT CTTGAATAAAAGAAAAACCT TCAGAACTGT TATTTTTAAA AAAATCTTCA AATATCCGTA GAACTTATTT5281 GTGAAACAAA TTAAGAATTA GTATCCATGA GAAAAATATA GAACAATTTT CCTAATTTAGCACTTTGTTT AATTCTTAAT CATAGGTACT CTTTTTATAT CTTGTTAAAA GGATTAAATC5341 TTTGAAAATC TGGGATTGAA GATGTGTGTC AAGAGATGTT GGTGGCAAGA ACATTTTTTTAAACTTTTAG ACCCTAACTT CTACACACAG TTCTCTACAA CCACCGTTCT TGTAAAAAAA5401 TTCAAGAACT TATAAAAATG CAACAAAACA AACCATTTAA TACATTTTGG TCAAAATCAAAAGTTCTTGA ATATTTTTAC GTTGTTTTGT TTGGTAAATT ATGTAAAACC AGTTTTAGTT5461 TAATGTATTT TATTTTATGC TCCAAGGAGC ATAAAATTGG GGACTGGGCA AGAGAAACTGATTACATAAA ATAAAATACG AGGTTCCTCG TATTTTAACC CCTGACCCGT TCTCTTTGAC5521 ACACCCTGGT AAATTACCAA GAGATAAGTA CACAGTTCTA TGTAGAGAAA ATAAGCATAGTGTGGGACCA TTTAATGGTT CTCTATTCAT GTGTCAAGAT ACATCTCTTT TATTCGTATC5581 TGTATGATCT CTAAAATTAT GTGAGACAAA GGAGAGATGA CATTAGGCAT GTGGGGATGAACATACTAGA GATTTTAATA CACTCTGTTT CCTCTCTACT GTAATCCGTA CACCCCTACT5641 AGACTGAGTA GAGAAGAAAC AATCTAATCA GTCCAAGAAA ACATCTCGAT CAGTGGAACATCTGACTCAT CTCTTCTTTG TTAGATTAGT CAGGTTCTTT TGTAGAGCTA GTCACCTTGT5701 AATAGAAGAA ATGCTAAAAT GAAACAGAAG TCTTACTGGA AATAAAAGAT ATGCATAAGATTATCTTCTT TACGATTTTA CTTTGTCTTC AGAATGACCT TTATTTTCTA TACGTATTCT5761 CAAAAATTCA TGAAAATCAC TTAGTTTAGC AGAGAAAAGA TAAAAATAAA GTATGACCTTGTTTTTAAGT ACTTTTAGTG AATCAAATCG TCTCTTTTCT ATTTTTATTT CATACTGGAA5821 CTTCATATAC ATTGTTTGAT CATATGCACC TCAATAAAAC TGAGTCTCCA ACAGAAATGAGAAGTATATG TAACAAACTA GTATACGTGG AGTTATTTTG ACTCAGAGGT TGTCTTTACT5881 AACATTAATA TTTTGTTCAC TGCTCTAATC CCAGAATCTA AGCGATATCT GGCAATAAAATTGTAATTAT AAAACAAGTG ACGAGATTAG GGTCTTAGAT TCGCTATAGA CCGTTATTTT5941 ATAATAAATA TATATTTTTT AATAAATGAA TCAACCACTT AATTTTTCTG TAAATATCTGTATTATTTAT ATATAAAAAA TTATTTACTT AGTTGGTGAA TTAAAAAGAC ATTTATAGAC6001 TAACTTCTCT TCTGTCTTTC CAAAAACACT CATAAGTACT GTGAATGAGA TGAAAAAGAGATTGAAGAGA AGACAGAAAG GTTTTTGTGA GTATTCATGA CACTTACTCT ACTTTTTCTC6061 TGAAGTAGGA TATAGGCTGT TAGCAGAAAA CATCTGAATG GCTGGCAGTG AAACATTAACACTTCATCCT ATATCCGACA ATCGTCTTTT GTAGACTTAC CGACCGTCAC TTTGTAATTG6121 TTGAAATGTA AGATTAATGA GTAATAGTAA ATTTTAACCT TGGCCATATG ATAAAATGTTAACTTTACAT TCTAATTACT CATTATCATT TAAAATTGGA ACCGGTATAC TATTTTACAA6181 CATTAATATT TTTCTAGAAT ACAGGGCTTT TTGTTTTTGC CATGAGGTTT GCAGGATCTTGTAATTATAA AAAGATCTTA TGTCCCGAAA AACAAAAACG GTACTCCAAA CGTCCTAGAA6241 GGTTCCCTGA CCAGGGATCA AACCTGCACT CCCCTGGAAG CATGGAGTCT TGGACATTTGCCAAGGGACT GGTCCCTAGT TTGGACGTGA GGGGACCTTC GTACCTCAGA ACCTGTAAAC6301 TATTATACAC TATCTTTGGT TCCTTTTAAA GGGAAGTAAT TTTACTTAAA TAAGAAAATAATAATATGTG ATAGAAACCA AGGAAAATTT CCCTTCATTA AAATGAATTT ATTCTTTTAT6361 GATTGACAAG TAATACGCTG TTTCCTCATC TTCCCATTCA CAGGAATCGA GAGCCATGAACTAACTGTTC ATTATGCGAC AAAGGAGTAG AAGGGTAAGT GTCCTTAGCT CTCGGTACTT6421 GGTCCTCATC CTTGCCTGTC TGGTGGCTCT GGCCATTGCG ATCGCCCAGGAGTAG GAACGGACAG ACCACCGAGA CCGGTAACGC TAGCG
3.根据权利要求1所述的关中奶山羊酪蛋白基因启动子表达载体，其特征在于将关中奶山羊β-酪蛋白基因启动区域序列全长6465bp插入Clontech公司的商售质粒pcDNA3.1(-)得到的关中奶山羊酪蛋白基因启动子表达载体全长为11838bp的序列如下1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTGCACTCT CAGTACAATC TGCTCTGATGCTGCCTAGCC CTCTAGAGGG CTAGGGGATA CCACGTGAGA GTCATGTTAG ACGAGACTAC61 CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCGGGCGTATCAA TTCGGTCATA GACGAGGGAC GAACACACAA CCTCCAGCGA CTCATCACGC121 CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGCGCTCGTTTTA AATTCGATGT TGTTCCGTTC CGAACTGGCT GTTAACGTAC TTCTTAGACG181 TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATTAATCCCAATC CGCAAAACGC GACGAAGCGC TACATGCCCG GTCTATATGC GCAACTGTAA241 GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATACTAATAACTG ATCAATAATT ATCATTAGTT AATGCCCCAG TAATCAAGTA TCGGGTATAT301 TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACCACCTCAAGGC GCAATGTATT GAATGCCATT TACCGGGCGG ACCGACTGGC GGGTTGCTGG361 CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCCGGGCGGGTAA CTGCAGTTAT TACTGCATAC AAGGGTATCA TTGCGGTTAT CCCTGAAAGG421 ATTGACGTCA ATGGGTGGAG TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGTTAACTGCAGT TACCCACCTC ATAAATGCCA TTTGACGGGT GAACCGTCAT GTAGTTCACA481 ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATTTAGTATACGG TTCATGCGGG GGATAACTGC AGTTACTGCC ATTTACCGGG CGGACCGTAA541 ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCATACGGGTCAT GTACTGGAAT ACCCTGAAAG GATGAACCGT CATGTAGATG CATAATCAGT601 TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTGAGCGATAATG GTACCACTAC GCCAAAACCG TCATGTAGTT ACCCGCACCT ATCGCCAAAC661 ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACCTGAGTGCCCC TAAAGGTTCA GAGGTGGGGT AACTGCAGTT ACCCTCAAAC AAAACCGTGG721 AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCGTTTTAGTTGC CCTGAAAGGT TTTACAGCAT TGTTGAGGCG GGGTAACTGC GTTTACCCGC781 GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCACATCCGCACA TGCCACCCTC CAGATATATT CGTCTCGAGA GACCGATTGA TCTCTTGGGT841 CTGCTTACTG GCTTATCGAA ATTAATACGA CTCACTATAG GGAGACCCAA GCTGGCTAGCGACGAATGAC CGAATAGCTT TAATTATGCT GAGTGATATC CCTCTGGGTT CGACCGATCG901 GTTTAAACGG GCCCTCTAGA CAGATGATTT TGCAACCCCC TGCCTCAGGA GACACTGGGACAAATTTGCC CGGGAGATCT GTCTACTAAA ACGTTGGGGG ACGGAGTCCT CTGTGACCCT961 AATTTCCTGA GACATTTTTG ATTCCAAAAG CTGTGCAGTT GGTGCTTCTA CCATCTTCGTTTAAAGGACT CTGTAAAAAC TAAGGTTTTC GACACGTCAA CCACGAAGAT GGTAGAAGCA1021 GGTAGAGGTC AAGGATGCTG CTAAACATTC TACAACACAT TAAGAAAACC CCCACAACAACCATCTCCAG TTCCTACGAC GATTTGTAAG ATGTTGTGTA ATTCTTTTGG GGGTGTTGTT1081 AGAATTCTTC CGCCAAAAAT ATCAATAATA TGAAGGTTGA AAAATACTGG TCTAGCATGTTCTTAAGAAG GCGGTTTTTA TAGTTATTAT ACTTCCAACT TTTTATGACC AGATCGTACA1141 AGTATGTGCT CAATAGCAAG GAGAGAAAAG AAAGCCTTCC TCACTGATTA ATGCAAAGAATCATACACGA GTTATCGTTC CTCTCTTTTC TTTCGGAAGG AGTGACTAAT TACGTTTCTT1201 ATAGAGGAAA ACAATAGAAT GGGAAAGACT AGAGAGCTCT TCAAGCAAAT TAGAGATATCTATCTCCTTT TGTTATCTTA CCCTTTCTGA TCTCTCGAGA AGTTCGTTTA ATCTCTATAG1261 AAGGGAACAT TTCACGCAAA GATGGGCACA ATAAAGGACA GAAATTTTAT GGAGGAGTTGTTCCCTTGTA AAGTGCGTTT CTACCCGTGT TATTTCCTGT CTTTAAAATA CCTCCTCAAC1321 CTGATGGAGA GGGAGGCCTG GCGTGCTGCG ATTCCTGGGG TCGCAAAGAG TCGGACACAAGACTACCTCT CCCTCCGGAC CGCACGACGC TAAGGACCCC AGCGTTTCTC AGCCTGTGTT1381 CTGAGCGACT GAATTGAACT GAACTGAACT GGACAAAGCA GAAGATATTA AGAAGAGGTGGACTCGCTGA CTTAACTTGA CTTGACTTGA CCTGTTTCGT CTTCTATAAT TCTTCTCCAC1441 GTAAGAATAC ACAGAAGAAC AATATAAAAA AGATCTTCAT GACCCAGATA ACCACGATGACATTCTTATG TGTCTTCTTG TTATATTTTT TCTAGAAGTA CTGGGTCTAT TGGTGCTACT1501 TGTGATCACT CACCTAGAGC CAGACACCCT GGAATGCAAA GTCAAACGGC CTTAGAAAGCACACTAGTGA GTGGATCTCG GTCTGTGGGA CCTTACGTTT CAGTTTGCCG GAATCTTTCG1561 CTCACTATGA ACAAAGCTAG TGGAGGTAAT GGAATTCCAG TTGAGCTATT TCAAATCTTAGAGTGATACT TGTTTCGATC ACCTCCATTA CCTTAAGGTC AACTCGATAA AGTTTAGAAT1621 AAAGGTGATG CTGTGAAAGT GCTGCACTCA ATATGTCAGC AAATTTGGAA AACTCAGCAGTTTCCACTAC GACACTTTCA CGACGTGAGT TATACAGTCG TTTAAACCTT TTGAGTCGTC1681 TGGCCACAGG ACTGCCACAA TCCCAAAGAA AAGCAATGAC AAAGAATGTT CAAACACCCAACCGGTGTCC TGACGGTGTT AGGGTTTCTT TTCGTTACTG TTTCTTACAA GTTTGTGGGT1741 CATGATTGCA CTCATCTCAC ATGCTAGCAA AATAACTCTC AAAATTCTCC AAGCCAGGCTGTACTAACGT GAGTAGAGTG TACGATCGTT TTATTGAGAG TTTTAAGAGG TTCGGTCCGA1801 CCAACAGTAC GTGGACCATG AACTTCCAGA TGTTCAAGCT GGATTTAGAA AAGGCAGAGGGGTTGTCATG CACCTGGTAC TTGAAGGTCT ACAAGTTCGA CCTAAATCTT TTCCGTCTCC1861 AACCAGAGAT CAAATTGCCA ACATCCATTG GATCATCAAA AAAGCACGAG AGTTCCAGAATTGGTCTCTA GTTTAACGGT TGTAGGTAAC CTAGTAGTTT TTTCGTGCTC TCAAGGTCTT1921 AAACATCTGC TTTATTGACT ACGCTAAAGC CTTTGATTGT GTGGATCACA ATAAACTGTGTTTGTAGACG AAATAACTGA TGCGATTTCG GAAACTAACA CACCTAGTGT TATTTGACAC1981 GAAAATTCTT CAAGAGATGG GAATACCAGA CCACTTTACC TGCCTCCTGA GAAATCTGTACTTTTAAGAA GTTCTCTACC CTTATGGTCT GGTGAAATGG ACGGAGGACT CTTTAGACAT2041 TACAGGTCCA GAAGCAGCAG TTAGAACTGG ACATGGAACA ACAGACTGGT TCCAAACTGCATGTCCAGGT CTTCGTCGTC AATCTTGACC TGTACCTTGT TGTCTGACCA AGGTTTGACG2101 GAAAGGGGTA CATCAAGGAA TATTCATTGG AAGGATTGAT GCTGAAGCTG AAACTCCTATCTTTCCCCAT GTAGTTCCTT ATAAGTAACC TTCCTAACTA CGACTTCGAC TTTGAGGATA2161 ACTTTGGCCA CCTAATGTGA AGATCTGACT CATTGGAAAA GACTCCAATG CTGGGAAAGATGAAACCGGT GGATTACACT TCTAGACTGA GTAACCTTTT CTGAGGTTAC GACCCTTTCT2221 TTGAAGGCAG GAGAAGAGGA TGACAGAGGA TGAGATGGTT GGATGGGATC ACTGACTCAAAACTTCCGTC CTCTTCTCCT ACTGTCTCCT ACTCTACCAA CCTACCCTAG TGACTGAGTT2281 TGGACATGAG TTTGAGTAAG CTCCAGGGGT TGGTGGTGGA CAGGAAAGCC TGGCGTGCTGACCTGTACTC AAACTCATTC GAGGTCCCCA ACCACCACCT GTCCTTTCGG ACCGCACGAC2341 CAGTCCACAA GGTCACAAAG ATTCGGACAT GACTGAGTGA CTGAACTGAT ACTGATGTGCGTCAGGTGTT CCAGTGTTTC TAAGCCTGTA CTGACTCACT GACTTGACTA TGACTACACG2401 TCAACAAATG TATCTTGAAC TTGTGTGAAG TTCTATGGTC ACATGTAAAG GAAGAATAATAGTTGTTTAC ATAGAACTTG AACACACTTC AAGATACCAG TGTACATTTC CTTCTTATTA2461 CAGGATTAGC TGTGTGTCTT AGGAATCAGG GTTCTGAGTT TTATGTGTTC ATAGTATCTGGTCCTAATCG ACACACAGAA TCCTTAGTCC CAAGACTCAA AATACACAAG TATCATAGAC2521 CTGGTTCACA AAACATTTTT CTTATTCTCT GGTTCTTGAT TTACTTTATA AAGTAATCTTGACCAAGTGT TTTGTAAAAA GAATAAGAGA CCAAGAACTA AATGAAATAT TTCATTAGAA2581 AATAGTTATA CTTCACATAG ATACGAAATT ATTATATTTG GATAATCTCA TGGAAAGGATTTATCAATAT GAAGTGTATC TATGCTTTAA TAATATAAAC CTATTAGAGT ACCTTTCCTA2641 TAAATACTCC ATCTATTACG AGTAATGCTG AACTATCTAC TCCTACCTAA TAATTTGTCAATTTATGAGG TAGATAATGC TCATTACGAC TTGATAGATG AGGATGGATT ATTAAACAGT2701 GAATTCACTA ATTCTGTGTT ATATTGTTTC TAAATCTGAA TCATTATATG AATCCTCAGTCTTAAGTGAT TAAGACACAA TATAACAAAG ATTTAGACTT AGTAATATAC TTAGGAGTCA2761 ATTTTGTTTT CCTTCCTCTA TATTTTGGAA TTTATTAAAC AGTGCTTCAA ATAATTTTTATAAAACAAAA GGAAGGAGAT ATAAAACCTT AAATAATTTG TCACGAAGTT TATTAAAAAT2821 GGAAACTGAA GTTTTTAGTA ACAGCTCTAT CTCTAAATAG CTTTAGTATC TTGAAAAAGTCCTTTGACTT CAAAAATCAT TGTCGAGATA GAGATTTATC GAAATCATAG AACTTTTTCA2881 AATACAAATT CTCACATCCT TAATTTCCTC TTCTCTAAAA TATCTTTAAA ATATTCTATGTTATGTTTAA GAGTGTAGGA ATTAAAGGAG AAGAGATTTT ATAGAAATTT TATAAGATAC2941 AATGATATCT CTTAATATTT ATTTTTTTGG CAATCCAACA CAGCTTATGG GATCTTAGTTTTACTATAGA GAATTATAAA TAAAAAAACC GTTAGGTTGT GTCGAATACC CTAGAATCAA3001 CCCCAGTGAG GGATTATATC CATGCCAACT GCAGTGAAAG TACAAAATCC TAAACTGGACGGGGTCACTC CCTAATATAG GTACGGTTGA CGTCACTTTC ATGTTTTAGG ATTTGACCTG3061 TCACCAGGGA TTTCCCAATA TCTCCTCTAG TTCTTATTTC TGAATATTTT TGGTCCCTTTAGTGGTCCCT AAAGGGTTAT AGAGGAGATC AAGAATAAAG ACTTATAAAA ACCAGGGAAA3121 ATTGTACTCT TCATCCAACT TTTCTATTGA TTTCTTTCTT GAGGTTATTA TTTACTTGGTTAACATGAGA AGTAGGTTGA AAAGATAACT AAAGAAAGAA CTCCAATAAT AAATGAACCA3181 TTCAGTTAGA AATATATGCA AATCTCAGGA CTGCATATTT CAGATTCATT GGCCAATATGAAGTCAATCT TTATATACGT TTAGAGTCCT GACGTATAAA GTCTAAGTAA CCGGTTATAC3241 GGAAAAAACC TTTGGCTGAA CAAATCATGC TTATAAAAAA TAGTACTAGA GCATCCTACTCCTTTTTTGG AAACCGACTT GTTTAGTACG AATATTTTTT ATCATGATCT CGTAGGATGA3301 TTGACTATAT CTTGCTCCTC ATTCAGGGTT ATCTAATACA ATTTCCCCAC ATGAAATTCTAACTGATATA GAACGAGGAG TAAGTCCCAA TAGATTATGT TAAAGGGGTG TACTTTAAGA3361 TTTGCATTAT AAAAATGGAA GCTCTTAGGT AACATTGCAA AAATTCGAGT TGCTCATATGAAACGTAATA TTTTTACCTT CGAGAATCCA TTGTAACGTT TTTAAGCTCA ACGAGTATAC3421 GCACTTTGCT TCTTACTGGT CATTGTGTTC TGAGGCTTAC CTGGACAGGT GGTACCTGATCGTGAAACGA AGAATGACCA GTAACACAAG ACTCCGAATG GACCTGTCCA CCATGGACTA3481 GTCATCTTAA ATTGCTGGCT TTTTGATTTT CCATTGGACA AGCTTCTTTC TTTAGTATATCAGTAGAATT TAACGACCGA AAAACTAAAA GGTAACCTGT TCGAAGAAAG AAATCATATA3541 TGTTAAGGAT TTCCTTGATC AAGATTTTAC CTACTTTTCT GGTCCAATTG GTGAGAGACAACAATTCCTA AAGGAACTAG TTCTAAAATG GATGAAAAGA CCAGGTTAAC CACTCTCTGT3601 GTCATAAGGA AATGCTGTGT TTATTGCACA ATATGTAAAG CATCTTCCTG AGAAAATAAACAGTATTCCT TTACGACACA AATAACGTGT TATACATTTC GTAGAAGGAC TCTTTTATTT3661 AGGGAAATGT TGAATGGGAA GGATATGCTT TCTTTTGTAT TCCTTTTCTG AGAAATCAAATCCCTTTACA ACTTACCCTT CCTATACGAA AGAAAACATA AGGAAAAGAC TCTTTAGTTT3721 CTTTTTCACC TGTGGCCTTG GCCACCAAAA GCTAACAAAT AAAGGCATAT GAAGTAGCCAGAAAAAGTGG ACACCGGAAC CGGTGGTTTT CGATTGTTTA TTTCCGTATA CTTCATCGGT3781 AGGCCTTTTC TAGTTATATC TATAACACTG AGTTCATTTC ATCATTTATT TTCCTGACTTTCCGGAAAAG ATCAATATAG ATATTGTGAC TCAAGTAAAG TAGTAAATAA AAGGACTGAA3841 CCTCCTGGGT CCATATGAGC AGTCTTAGAA TGAATATTAG CTGAATAATC CAAATACATAGGAGGACCCA GGTATACTCG TCAGAATCTT ACTTATAATC GACTTATTAG GTTTATGTAT3901 GTAGATGTTG ATTTGGGTTT TCTAAGCAAT CCAAGACTTG TATGACAGTA AGATGTATTACATCTACAAC TAAACCCAAA AGATTCGTTA GGTTCTGAAC ATACTGTCAT TCTACATAAT3961 CCATCCAACA CACATCTCAG CATGATATAA ATGCAAGGTA TATTGTGAAG AAAAATTTTTGGTAGGTTGT GTGTAGAGTC GTACTATATT TACGTTCCAT ATAACACTTC TTTTTAAAAA4021 AATTATGTCA AAGTGCTTAC TTTAGAAGGT CATCTATCTG TCCCAAAGCT GTGAATATATTTAATACAGT TTCACGAATG AAATCTTCCA GTAGATAGAC AGGGTTTCGA CACTTATATA4081 ATATTGAAGG TAATGAATAG ATGAAGCTAA CCTTGTAAAA ATGAGTAGTG TGAAATACAATATAACTTCC ATTACTTATC TACTTCGATT GGAACATTTT TACTCATCAC ACTTTATGTT4141 CTACAATTAT GAACATCTGT CACTAAAGAG GCAAAGAAAC TTGAAGATTG CTTTTGCAAAGATGTTAATA CTTGTAGACA GTGATTTCTC CGTTTCTTTG AACTTCTAAC GAAAACGTTT4201 TGGGCTCCTA TTAATAAAAA GTACTTTTGA GGTCTGGCTC AGACTCTATT GTAGTACTTAACCCGAGGAT AATTATTTTT CATGAAAACT CCAGACCGAG TCTGAGATAA CATCATGAAT4261 GGGTAAGACC CTCCTCCTGT ATGGGCTTTC ATTTTCTTTC TTGCTTCCCT CATTTGCCCTCCCATTCTGG GAGGAGGACA TACCCGAAAG TAAAAGAAAG AACGAAGGGA GTAAACGGGA4321 TCCATGAATA CTAGCTGATA AACATTGACT ATAAAAGATA TGAGGCCAAA CTTGAGCTGTAGGTACTTAT GATCGACTAT TTGTAACTGA TATTTTCTAT ACTCCGGTTT GAACTCGACA4381 CCCATTTTAA TAAATCTGTA TAAATAATAT TTGTTCTACA AAAGTATTAT CTAAATAAATGGGTAAAATT ATTTAGACAT ATTTATTATA AACAAGATGT TTTCATAATA GATTTATTTA4441 GTTACTTTCT GTCTTAAAAT CCCTCAACAA ATCCCCACTA TCTAGAGAAT AAGATTGACACAATGAAAGA CAGAATTTTA GGGAGTTGTT TAGGGGTGAT AGATCTCTTA TTCTAACTGT4501 TTCCCTGGAA TCACAGCATG CTTTGTCTGC CATTATCTGA CCCCTTTCTC TTTCTCTCTTAAGGGACCTT AGTGTCGTAC GAAACAGACG GTAATAGACT GGGGAAAGAG AAAGAGAGAA4561 CTCACCTCCA TCTACTCCTT TTTCCTTGCA ATTCATGACC CAGATTCACT GTTTGATTTGGAGTGGAGGT AGATGAGGAA AAAGGAACGT TAAGTACTGG GTCTAAGTGA CAAACTAAAC4621 GCTTGCATGT GTGTGTGCTG AGTTGCGTCT GACTGTTATC AACCCCATGA ATGATAGTCCCGAACGTACA CACACACGAC TCAACGCAGA CTGACAATAG TTGGGGTACT TACTATCAGG4681 ACCAGGCTCT ACTGTCCATG AAATTTTCCA GTCAAGAATA CTGGAGTGGA TTGCATTTCCTGGTCCGAGA TGACAGGTAC TTTAAAAGGT CAGTTCTTAT GACCTCACCT AACGTAAAGG4741 TACTCCATTT GATTAATTTA GTGACTTTTA AATTTCTTTT TCCATATTCG GGAGCCTATTATGAGGTAAA CTAATTAAAT CACTGAAAAT TTAAAGAAAA AGGTATAAGC CCTCGGATAA4801 CTTCCTTTTT AGTCTATACT CTCTTCACTC TTCAGGTCTA AGGTATCATC GTGTGCTTGTGAAGGAAAAA TCAGATATGA GAGAAGTGAG AAGTCCAGAT TCCATAGTAG CACACGAACA4861 TAGCTTGTTA CTTTCTCCAT TATAGCTTAA GCACTAACAA CTGTTCAGGT TGGCATGAAAATCGAACAAT GAAAGAGGTA ATATCGAATT CGTGATTGTT GACAAGTCCA ACCGTACTTT4921 TTGTGTTCTT TGTGTGGCCT GTATATTTCT GTTGTGTATT AGAATTTACC CCAAGATCTCAACACAAGAA ACACACCGGA CATATAAAGA CAACACATAA TCTTAAATGG GGTTCTAGAG4981 AAAGACCCAC TGAATACTAA AGAGACCTCA TTGTGGTTAC AATAATTTGG GGACTGGGCCTTTCTGGGTG ACTTATGATT TCTCTGGAGT AACACCAATG TTATTAAACC CCTGACCCGG5041 AAAACTACCG TGCATCCCAG CCAAGATCTG TAGCTACTGG ACAATTTCAT TTCCTTTATCTTTTGATGGC ACGTAGGGTC GGTTCTAGAC ATCGATGACC TGTTAAAGTA AAGGAAATAG5101 AGATTGTGAG TTATTCCTGT TAAAATGCTC CCCAGAATTT CTGGGGACAG AAAAATAGGATCTAACACTC AATAAGGACA ATTTTACGAG GGGTCTTAAA GACCCCTGTC TTTTTATCCT5161 AGAATTCATT TCCTAATCAT GCAGATTTCT AGGAATTCAA ATCCACTGTT GGTTTTATTTTCTTAAGTAA AGGATTAGTA CGTCTAAAGA TCCTTAAGTT TAGGTGACAA CCAAAATAAA5221 CAAACCACAA AATTAGCATG CCATTAAATA CTATATATAA ACAGCCACTA AATCAGATCAGTTTGGTGTT TTAATCGTAC GGTAATTTAT GATATATATT TGTCGGTGAT TTAGTCTAGT5281 TTATCCATTC AGCTTCTCCT TCACTTCTTC TCCTCTACTT TGGAAAAAAG GTAAGAATCTAATAGGTAAG TCGAAGAGGA AGTGAAGAAG AGGAGATGAA ACCTTTTTTC CATTCTTAGA5341 CAGATATAAT TTCAGGTGTA TCTGCTACTC ATCTTTATTT TGGACTAGGT TAAAATGTAGGTCTATATTA AAGTCCACAT AGACGATGAG TAGAAATAAA ACCTGATCCA ATTTTACATC5401 AAAGAACATA ATTGCTTAAA ATAGATCTTA AAAATAAGGG TGTTTAAGAT AAGGTTTACATTTCTTGTAT TAACGAATTT TATCTAGAAT TTTTATTCCC ACAAATTCTA TTCCAAATGT5461 CTATTTTCAG CAGATATGTT AAAAAATAGA AGTGACTATA AAGACTTGAT AAAAATTATAGATAAAAGTC GTCTATACAA TTTTTTATCT TCACTGATAT TTCTGAACTA TTTTTAATAT5521 GTGACTGCAA ATGTTTTAGG AATATAATAA GATATAATAA CGGTGGTTGC TATTTTCTTTCACTGACGTT TACAAAATCC TTATATTATT CTATATTATT GCCACCAACG ATAAAAGAAA5581 AGCACAAGAC TAGTTAACAG GCTGTATTAA AAGATCTTTT CTTGAATTAA ATATTTTCAATCGTGTTCTG ATCAATTGTC CGACATAATT TTCTAGAAAA GAACTTAATT TATAAAAGTT5641 TTTGATTAAA CCTACCTCAG CCATAAAGGC AAGCACATTT CATTTATACT ATGGGGATTTAAACTAATTT GGATGGAGTC GGTATTTCCG TTCGTGTAAA GTAAATATGA TACCCCTAAA5701 GAATAATTAT TACTGAAGAA GCTCTACCAA CAAAAAGTTT ATAGAGCTAT CATATTTAGTCTTATTAATA ATGACTTCTT CGAGATGGTT GTTTTTCAAA TATCTCGATA GTATAAATCA5761 CAAGAGATAA AGAGGGTTGT TAGGATATAT ATGCTATTTG AAAGGTATTT ATAAAAGAAGGTTCTCTATT TCTCCCAACA ATCCTATATA TACGATAAAC TTTCCATAAA TATTTTCTTC5821 AGTATATTTA TCAAAATTTC TCAGAACATC CAAATTTCAA GTTTATCATT TATCTTACAATCATATAAAT AGTTTTAAAG AGTCTTGTAG GTTTAAAGTT CAAATAGTAA ATAGAATGTT5881 TATTTCAAAA ATATTAAAAT AGATACTGAA ATACAGAAGT AAATTAAAGA GAAAGTATTTATAAAGTTTT TATAATTTTA TCTATGACTT TATGTCTTCA TTTAATTTCT CTTTCATAAA5941 TACTTGGTAA AAAAATTCTA GGTTGGACAG AGAGTGCCAG GAAACAAAAA CAATGAAAAAATGAACCATT TTTTTAAGAT CCAACCTGTC TCTCACGGTC CTTTGTTTTT GTTACTTTTT6001 TGTGACCTGA CAGGAATTAT AGCTCAAAGT ATAGTAGTAA GTAATGAAAT GGCTTAAAAAACACTGGACT GTCCTTAATA TCGAGTTTCA TATCATCATT CATTACTTTA CCGAATTTTT6061 TTGGTATATA AAATGCTAGT TATAAAATAA ACAAAATGCA ATAATATCCT CCCTACATGTAACCATATAT TTTACGATCA ATATTTTATT TGTTTTACGT TATTATAGGA GGGATGTACA6121 AATGAATTCT AGGTATTATG CTCTTTTTGG AAGTCTTGAC AATAAAAATT TTTTTAGAAGTTACTTAAGA TCCATAATAC GAGAAAAACC TTCAGAACTG TTATTTTTAA AAAAATCTTC6181 TTTATAGGCA TCTTGAATAA AGTGAAACAA ATTAAGAATT AGTATCCATG AGAAAAATATAAATATCCGT AGAACTTATT TCACTTTGTT TAATTCTTAA TCATAGGTAC TCTTTTTATA6241 AGAACAATTT TCCTAATTTA GTTTGAAAAT CTGGGATTGA AGATGTGTGT CAAGAGATGTTCTTGTTAAA AGGATTAAAT CAAACTTTTA GACCCTAACT TCTACACACA GTTCTCTACA6301 TGGTGGCAAG AACATTTTTT TTTCAAGAAC TTATAAAAAT GCAACAAAAC AAACCATTTAACCACCGTTC TTGTAAAAAA AAAGTTCTTG AATATTTTTA CGTTGTTTTG TTTGGTAAAT6361 ATACATTTTG GTCAAAATCA ATAATGTATT TTATTTTATG CTCCAAGGAG CATAAAATTGTATGTAAAAC CAGTTTTAGT TATTACATAA AATAAAATAC GAGGTTCCTC GTATTTTAAC6421 GGGACTGGGC AAGAGAAACT GACACCCTGG TAAATTACCA AGAGATAAGT ACACAGTTCTCCCTGACCCG TTCTCTTTGA CTGTGGGACC ATTTAATGGT TCTCTATTCA TGTGTCAAGA6481 ATGTAGAGAA AATAAGCATA GTGTATGATC TCTAAAATTA TGTGAGACAA AGGAGAGATGTACATCTCTT TTATTCGTAT CACATACTAG AGATTTTAAT ACACTCTGTT TCCTCTCTAC6541 ACATTAGGCA TGTGGGGATG AAGACTGAGT AGAGAAGAAA CAATCTAATC AGTCCAAGAATGTAATCCGT ACACCCCTAC TTCTGACTCA TCTCTTCTTT GTTAGATTAG TCAGGTTCTT6601 AACATCTCGA TCAGTGGAAC AAATAGAAGA AATGCTAAAA TGAAACAGAA GTCTTACTGGTTGTAGAGCT AGTCACCTTG TTTATCTTCT TTACGATTTT ACTTTGTCTT CAGAATGACC6661 AAATAAAAGA TATGCATAAG ACAAAAATTC ATGAAAATCA CTTAGTTTAG CAGAGAAAAGTTTATTTTCT ATACGTATTC TGTTTTTAAG TACTTTTAGT GAATCAAATC GTCTCTTTTC6721 ATAAAAATAA AGTATGACCT TCTTCATATA CATTGTTTGA TCATATGCAC CTCAATAAAATATTTTTATT TCATACTGGA AGAAGTATAT GTAACAAACT AGTATACGTG GAGTTATTTT6781 CTGAGTCTCC AACAGAAATG AAACATTAAT ATTTTGTTCA CTGCTCTAAT CCCAGAATCTGACTCAGAGG TTGTCTTTAC TTTGTAATTA TAAAACAAGT GACGAGATTA GGGTCTTAGA6841 AAGCGATATC TGGCAATAAA AATAATAAAT ATATATTTTT TAATAAATGA ATCAACCACTTTCGCTATAG ACCGTTATTT TTATTATTTA TATATAAAAA ATTATTTACT TAGTTGGTGA6901 TAATTTTTCT GTAAATATCT GTAACTTCTC TTCTGTCTTT CCAAAAACAC TCATAAGTACATTAAAAAGA CATTTATAGA CATTGAAGAG AAGACAGAAA GGTTTTTGTG AGTATTCATG6961 TGTGAATGAG ATGAAAAAGA GTGAAGTAGG ATATAGGCTG TTAGCAGAAA ACATCTGAATACACTTACTC TACTTTTTCT CACTTCATCC TATATCCGAC AATCGTCTTT TGTAGACTTA7021 GGCTGGCAGT GAAACATTAA CTTGAAATGT AAGATTAATG AGTAATAGTA AATTTTAACCCCGACCGTCA CTTTGTAATT GAACTTTACA TTCTAATTAC TCATTATCAT TTAAAATTGG7081 TTGGCCATAT GATAAAATGT TCATTAATAT TTTTCTAGAA TACAGGGCTT TTTGTTTTTGAACCGGTATA CTATTTTACA AGTAATTATA AAAAGATCTT ATGTCCCGAA AAACAAAAAC7141 CCATGAGGTT TGCAGGATCT TGGTTCCCTG ACCAGGGATC AAACCTGCAC TCCCCTGGAAGGTACTCCAA ACGTCCTAGA ACCAAGGGAC TGGTCCCTAG TTTGGACGTG AGGGGACCTT7201 GCATGGAGTC TTGGACATTT GTATTATACA CTATCTTTGG TTCCTTTTAA AGGGAAGTAACGTACCTCAG AACCTGTAAA CATAATATGT GATAGAAACC AAGGAAAATT TCCCTTCATT7261 TTTTACTTAA ATAAGAAAAT AGATTGACAA GTAATACGCT GTTTCCTCAT CTTCCCATTCAAAATGAATT TATTCTTTTA TCTAACTGTT CATTATGCGA CAAAGGAGTA GAAGGGTAAG7321 ACAGGAATCG AGAGCCATGA AGGTCCTCAT CCTTGCCTGT CTGGTGGCTC TGGCCATTGCTGTCCTTAGC TCTCGGTACT TCCAGGAGTA GGAACGGACA GACCACCGAG ACCGGTAACG7381 GATCGCGGAT CCGAGCTCGG TACCAAGCTT AAGTTTAAAC CCGCTGATCA GCCTCGACTGCTAGCGCCTA GGCTCGAGCC ATGGTTCGAA TTCAAATTTG GGCGACTAGT CGGAGCTGAC7441 TGCCTTCTAG TTGCCAGCCA TCTGTTGTTT GCCCCTCCCC CGTGCCTTCC TTGACCCTGGACGGAAGATC AACGGTCGGT AGACAACAAA CGGGGAGGGG GCACGGAAGG AACTGGGACC7501 AAGGTGCCAC TCCCACTGTC CTTTCCTAAT AAAATGAGGA AATTGCATCG CATTGTCTGATTCCACGGTG AGGGTGACAG GAAAGGATTA TTTTACTCCT TTAACGTAGC GTAACAGACT7561 GTAGGTGTCA TTCTATTCTG GGGGGTGGGG TGGGGCAGGA CAGCAAGGGG GAGGATTGGGCATCCACAGT AAGATAAGAC CCCCCACCCC ACCCCGTCCT GTCGTTCCCC CTCCTAACCC7621 AAGACAATAG CAGGCATGCT GGGGATGCGG TGGGCTCTAT GGCTTCTGAG GCGGAAAGAATTCTGTTATC GTCCGTACGA CCCCTACGCC ACCCGAGATA CCGAAGACTC CGCCTTTCTT7681 CCAGCTGGGG CTCTAGGGGG TATCCCCACG CGCCCTGTAG CGGCGCATTA AGCGCGGCGGGGTCGACCCC GAGATCCCCC ATAGGGGTGC GCGGGACATC GCCGCGTAAT TCGCGCCGCC7741 GTGTGGTGGT TACGCGCAGC GTGACCGCTA CACTTGCCAG CGCCCTAGCG CCCGCTCCTTCACACCACCA ATGCGCGTCG CACTGGCGAT GTGAACGGTC GCGGGATCGC GGGCGAGGAA7801 TCGCTTTCTT CCCTTCCTTT CTCGCCACGT TCGCCGGCTT TCCCCGTCAA GCTCTAAATCAGCGAAAGAA GGGAAGGAAA GAGCGGTGCA AGCGGCCGAA AGGGGCAGTT CGAGATTTAG7861 GGGGGCTCCC TTTAGGGTTC CGATTTAGTG CTTTACGGCA CCTCGACCCC AAAAAACTTGCCCCCGAGGG AAATCCCAAG GCTAAATCAC GAAATGCCGT GGAGCTGGGG TTTTTTGAAC7921 ATTAGGGTGA TGGTTCACGT AGTGGGCCAT CGCCCTGATA GACGGTTTTT CGCCCTTTGATAATCCCACT ACCAAGTGCA TCACCCGGTA GCGGGACTAT CTGCCAAAAA GCGGGAAACT7981 CGTTGGAGTC CACGTTCTTT AATAGTGGAC TCTTGTTCCA AACTGGAACA ACACTCAACCGCAACCTCAG GTGCAAGAAA TTATCACCTG AGAACAAGGT TTGACCTTGT TGTGAGTTGG8041 CTATCTCGGT CTATTCTTTT GATTTATAAG GGATTTTGCC GATTTCGGCC TATTGGTTAAGATAGAGCCA GATAAGAAAA CTAAATATTC CCTAAAACGG CTAAAGCCGG ATAACCAATT8101 AAAATGAGCT GATTTAACAA AAATTTAACG CGAATTAATT CTGTGGAATG TGTGTCAGTTTTTTACTCGA CTAAATTGTT TTTAAATTGC GCTTAATTAA GACACCTTAC ACACAGTCAA8161 AGGGTGTGGA AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAATCCCACACCT TTCAGGGGTC CGAGGGGTCG TCCGTCTTCA TACGTTTCGT ACGTAGAGTT8221 TTAGTCAGCA ACCAGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAGAATCAGTCGT TGGTCCACAC CTTTCAGGGG TCCGAGGGGT CGTCCGTCTT CATACGTTTC8281 CATGCATCTC AATTAGTCAG CAACCATAGT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCTGTACGTAGAG TTAATCAGTC GTTGGTATCA GGGCGGGGAT TGAGGCGGGT AGGGCGGGGA8341 AACTCCGCCC AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGCTTGAGGCGGG TCAAGGCGGG TAAGAGGCGG GGTACCGACT GATTAAAAAA AATAAATACG8401 AGAGGCCGAG GCCGCCTCTG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGGTCTCCGGCTC CGGCGGAGAC GGAGACTCGA TAAGGTCTTC ATCACTCCTC CGAAAAAACC8461 AGGCCTAGGC TTTTGCAAAA AGCTCCCGGG AGCTTGTATA TCCATTTTCG GATCTGATCATCCGGATCCG AAAACGTTTT TCGAGGGCCC TCGAACATAT AGGTAAAAGC CTAGACTAGT8521 AGAGACAGGA TGAGGATCGT TTCGCATGAT TGAACAAGAT GGATTGCACG CAGGTTCTCCTCTCTGTCCT ACTCCTAGCA AAGCGTACTA ACTTGTTCTA CCTAACGTGC GTCCAAGAGG8581 GGCCGCTTGG GTGGAGAGGC TATTCGGCTA TGACTGGGCA CAACAGACAA TCGGCTGCTCCCGGCGAACC CACCTCTCCG ATAAGCCGAT ACTGACCCGT GTTGTCTGTT AGCCGACGAG8641 TGATGCCGCC GTGTTCCGGC TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG TCAAGACCGAACTACGGCGG CACAAGGCCG ACAGTCGCGT CCCCGCGGGC CAAGAAAAAC AGTTCTGGCT8701 CCTGTCCGGT GCCCTGAATG AACTGCAGGA CGAGGCAGCG CGGCTATCGT GGCTGGCCACGGACAGGCCA CGGGACTTAC TTGACGTCCT GCTCCGTCGC GCCGATAGCA CCGACCGGTG8761 GACGGGCGTT CCTTGCGCAG CTGTGCTCGA CGTTGTCACT GAAGCGGGAA GGGACTGGCTCTGCCCGCAA GGAACGCGTC GACACGAGCT GCAACAGTGA CTTCGCCCTT CCCTGACCGA8821 GCTATTGGGC GAAGTGCCGG GGCAGGATCT CCTGTCATCT CACCTTGCTC CTGCCGAGAACGATAACCCG CTTCACGGCC CCGTCCTAGA GGACAGTAGA GTGGAACGAG GACGGCTCTT8881 AGTATCCATC ATGGCTGATG CAATGCGGCG GCTGCATACG CTTGATCCGG CTACCTGCCCTCATAGGTAG TACCGACTAC GTTACGCCGC CGACGTATGC GAACTAGGCC GATGGACGGG8941 ATTCGACCAC CAAGCGAAAC ATCGCATCGA GCGAGCACGT ACTCGGATGG AAGCCGGTCTTAAGCTGGTG GTTCGCTTTG TAGCGTAGCT CGCTCGTGCA TGAGCCTACC TTCGGCCAGA9001 TGTCGATCAG GATGATCTGG ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG AACTGTTCGCACAGCTAGTC CTACTAGACC TGCTTCTCGT AGTCCCCGAG CGCGGTCGGC TTGACAAGCG9061 CAGGCTCAAG GCGCGCATGC CCGACGGCGA GGATCTCGTC GTGACCCATG GCGATGCCTGGTCCGAGTTC CGCGCGTACG GGCTGCCGCT CCTAGAGCAG CACTGGGTAC CGCTACGGAC9121 CTTGCCGAAT ATCATGGTGG AAAATGGCCG CTTTTCTGGA TTCATCGACT GTGGCCGGCTGAACGGCTTA TAGTACCACC TTTTACCGGC GAAAAGACCT AAGTAGCTGA CACCGGCCGA9181 GGGTGTGGCG GACCGCTATC AGGACATAGC GTTGGCTACC CGTGATATTG CTGAAGAGCTCCCACACCGC CTGGCGATAG TCCTGTATCG CAACCGATGG GCACTATAAC GACTTCTCGA9241 TGGCGGCGAA TGGGCTGACC GCTTCCTCGT GCTTTACGGT ATCGCCGCTC CCGATTCGCAACCGCCGCTT ACCCGACTGG CGAAGGAGCA CGAAATGCCA TAGCGGCGAG GGCTAAGCGT9301 GCGCATCGCC TTCTATCGCC TTCTTGACGA GTTCTTCTGA GCGGGACTCT GGGGTTCGAACGCGTAGCGG AAGATAGCGG AAGAACTGCT CAAGAAGACT CGCCCTGAGA CCCCAAGCTT9361 ATGACCGACC AAGCGACGCC CAACCTGCCA TCACGAGATT TCGATTCCAC CGCCGCCTTCTACTGGCTGG TTCGCTGCGG GTTGGACGGT AGTGCTCTAA AGCTAAGGTG GCGGCGGAAG9421 TATGAAAGGT TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT CCTCCAGCGCATACTTTCCA ACCCGAAGCC TTAGCAAAAG GCCCTGCGGC CGACCTACTA GGAGGTCGCG9481 GGGGATCTCA TGCTGGAGTT CTTCGCCCAC CCCAACTTGT TTATTGCAGC TTATAATGGTCCCCTAGAGT ACGACCTCAA GAAGCGGGTG GGGTTGAACA AATAACGTCG AATATTACCA9541 TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCTATGTTTATTT CGTTATCGTA GTGTTTAAAG TGTTTATTTC GTAAAAAAAG TGACGTAAGA9601 AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATCATG TCTGTATACC GTCGACCTCTTCAACACCAA ACAGGTTTGA GTAGTTACAT AGAATAGTAC AGACATATGG CAGCTGGAGA9661 AGCTAGAGCT TGGCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAATTG TTATCCGCTCTCGATCTCGA ACCGCATTAG TACCAGTATC GACAAAGGAC ACACTTTAAC AATAGGCGAG9721 ACAATTCCAC ACAACATACG AGCCGGAAGC ATAAAGTGTA AAGCCTGGGG TGCCTAATGATGTTAAGGTG TGTTGTATGC TCGGCCTTCG TATTTCACAT TTCGGACCCC ACGGATTACT9781 GTGAGCTAAC TCACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTGCACTCGATTG AGTGTAATTA ACGCAACGCG AGTGACGGGC GAAAGGTCAG CCCTTTGGAC9841 TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGGAGCACGGTCG ACGTAATTAC TTAGCCGGTT GCGCGCCCCT CTCCGCCAAA CGCATAACCC9901 CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCGGCGAGAAGGC GAAGGAGCGA GTGACTGAGC GACGCGAGCC AGCAAGCCGA CGCCGCTCGC9961 GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA TAACGCAGGACATAGTCGAG TGAGTTTCCG CCATTATGCC AATAGGTGTC TTAGTCCCCT ATTGCGTCCT10021 AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC CGCGTTGCTGTTCTTGTACA CTCGTTTTCC GGTCGTTTTC CGGTCCTTGG CATTTTTCCG GCGCAACGAC10081 GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG CTCAAGTCAGCGCAAAAAGG TATCCGAGGC GGGGGGACTG CTCGTAGTGT TTTTAGCTGC GAGTTCAGTC10141 AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTCTCCACCGCTT TGGGCTGTCC TGATATTTCT ATGGTCCGCA AAGGGGGACC TTCGAGGGAG10201 GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCGCACGCGAGAG GACAAGGCTG GGACGGCGAA TGGCCTATGG ACAGGCGGAA AGAGGGAAGC10261 GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT GTAGGTCGTTCCTTCGCACC GCGAAAGAGT ATCGAGTGCG ACATCCATAG AGTCAAGCCA CATCCAGCAA10321 CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG CGCCTTATCCGCGAGGTTCG ACCCGACACA CGTGCTTGGG GGGCAAGTCG GGCTGGCGAC GCGGAATAGG10381 GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT GGCAGCAGCCCCATTGATAG CAGAACTCAG GTTGGGCCAT TCTGTGCTGA ATAGCGGTGA CCGTCGTCGG10441 ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGGTGACCATTGT CCTAATCGTC TCGCTCCATA CATCCGCCAC GATGTCTCAA GAACTTCACC10501 TGGCCTAACT ACGGCTACAC TAGAAGAACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCAACCGGATTGA TGCCGATGTG ATCTTCTTGT CATAAACCAT AGACGCGAGA CGACTTCGGT10561 GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC CGCTGGTAGCCAATGGAAGC CTTTTTCTCA ACCATCGAGA ACTAGGCCGT TTGTTTGGTG GCGACCATCG10621 GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCTCCAAAAAAAC AAACGTTCGT CGTCTAATGC GCGTCTTTTT TTCCTAGAGT TCTTCTAGGA10681 TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTGAACTAGAAAA GATGCCCCAG ACTGCGAGTC ACCTTGCTTT TGAGTGCAAT TCCCTAAAAC10741 GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTTCAGTACTCTA ATAGTTTTTC CTAGAAGTGG ATCTAGGAAA ATTTAATTTT TACTTCAAAA10801 AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGTTTTAGTTAGA TTTCATATAT ACTCATTTGA ACCAGACTGT CAATGGTTAC GAATTAGTCA10861 GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTCCTCCGTGGAT AGAGTCGCTA GACAGATAAA GCAAGTAGGT ATCAACGGAC TGAGGGGCAG10921 GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCGCACATCTATT GATGCTATGC CCTCCCGAAT GGTAGACCGG GGTCACGACG TTACTATGGC10981 CGAGACCCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCCGCTCTGGGTG CGAGTGGCCG AGGTCTAAAT AGTCGTTATT TGGTCGGTCG GCCTTCCCGG11041 GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGGCTCGCGTCTT CACCAGGACG TTGAAATAGG CGGAGGTAGG TCAGATAATT AACAACGGCC11101 GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACACTTCGATCTC ATTCATCAAG CGGTCAATTA TCAAACGCGT TGCAACAACG GTAACGATGT11161 GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGACCGTAGCACC ACAGTGCGAG CAGCAAACCA TACCGAAGTA AGTCGAGGCC AAGGGTTGCT11221 TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCTAGTTCCGCTC AATGTACTAG GGGGTACAAC ACGTTTTTTC GCCAATCGAG GAAGCCAGGA11281 CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTGGGCTAGCAAC AGTCTTCATT CAACCGGCGT CACAATAGTG AGTACCAATA CCGTCGTGAC11341 CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCAGTATTAAGAG AATGACAGTA CGGTAGGCAT TCTACGAAAA GACACTGACC ACTCATGAGT11401 ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATATGGTTCAGTA AGACTCTTAT CACATACGCC GCTGGCTCAA CGAGAACGGG CCGCAGTTAT11461 CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCTGCCCTATTAT GGCGCGGTGT ATCGTCTTGA AATTTTCACG AGTAGTAACC TTTTGCAAGA11521 TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACTAGCCCCGCTT TTGAGAGTTC CTAGAATGGC GACAACTCTA GGTCAAGCTA CATTGGGTGA11581 CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAAGCACGTGGGT TGACTAGAAG TCGTAGAAAA TGAAAGTGGT CGCAAAGACC CACTCGTTTT11641 ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTCTGTCCTTCCG TTTTACGGCG TTTTTTCCCT TATTCCCGCT GTGCCTTTAC AACTTATGAG11701 ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGATATGAGAAGG AAAAAGTTAT AATAACTTCG TAAATAGTCC CAATAACAGA GTACTCGCCT11761 TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGAATGTATAAAC TTACATAAAT CTTTTTATTT GTTTATCCCC AAGGCGCGTG TAAAGGGGCT11821 AAAGTGCCAC CTGACGTCTTTCACGGTG GACTGCAG
全文摘要
本发明涉及一种用关中奶山羊酪蛋白基因启动子区域构建的表达载体。特征是载体DNA序列中包含关中奶山羊乳腺组织中β－酪蛋白基因的启动子及其周围的调控序列以及β－酪蛋白基因的第一内含子、第一外显子和第二外显子及其信号肽6.5kb；将这段6.5kb长的序列插入商售的质粒载体中，构建得到含关中奶山羊β－酪蛋白基因启动子区域序列的重组质粒，DNA序列全长为6.5kb加上商售的质粒载体序列；实验结果充分说明所克隆的6.5kb DNA序列具有启动子和增强子活性。用本启动子构建的表达载体能够用于建立转基因山羊乳腺生物反应器，使用基因工程手段所构建的目的蛋白特异性地在山羊乳腺中表达、并分泌到山羊乳汁中，以供制备生产目的蛋白。
文档编号C12N15/63GK1661018SQ20041007316
公开日2005年8月31日 申请日期2004年10月11日 优先权日2004年10月11日
发明者陈苏民, 梁克明, 董德文 申请人:中国人民解放军第四军医大学