海州常山不同组织的荧光定量内参基因及其引物和应用的制作方法

本发明属于植物分子生物学领域，具体涉及海州常山不同组织的荧光定量内参基因及其引物和应用。技术背景海州常山(clerodendrumtrichotomumthunb.)又名臭梧桐，马鞭草科(verbenaceae)大青属(clerodendruml.)下落叶灌木或小乔木，海州常山叶片较大可入药，据报道叶片还可作为野菜食用，花美果艳，花萼紫红色，5裂几达基部，花期时间长且花量巨大。核果球形，熟时蓝紫色，有光泽，包藏于宿萼内，秋冬经久不落，是秋冬季良好的观果树种。海州常山不仅有药用的功能还兼具盐碱、耐干旱等优良特性，是一种值得开发的优良的园林绿化树种。现在关于海州常山组织中分子机理的研究还未曾有报道，据研究表明没有一个内参基因在所有实验条件下恒定表达，常用的内参基因在不同的植物及实验条件下会表现不稳定的形状，从而影响目的基因检测的准确性，所以要开发海州常山中优良的基因，必须确定稳定可靠的内参基因。技术实现要素：发明目的：针对现有技术中存在的不足，本发明目的在于提供海州常山不同组织的荧光定量内参基因，该基因能够满足实时荧光定量检测海州常转录表达水平的要求，提高了海州常山基因表达分析研究的稳定性、可靠性和效率。本发明的另一目的是提供一种上述内参基因的专用引物。本发明还有一目的是提供一种上述内参基因或专用引物的应用。技术方案：为了实现上述发明目的，本发明采用的技术方案为：海州常山不同组织的荧光定量内参基因，为actin、rpl、ran、ap-2和hsp70基因，其中，actin基因的基因序列如seqidno.1所示，rpl基因的基因序列如seqidno.3所示，ran基因的基因序列如seqidno.6所示，ap-2基因的基因序列如seqidno.13所示，hsp70基因的基因序列如seqidno.14所示。所述的海州常山不同组织的荧光定量内参基因的专用引物，rpl基因的引物序列如下：rpl正向引物5’-ccacaactaacgcctccacc-3’rpl反向引物5’-aaattcaccctccgactccc-3’。所述的海州常山不同组织的荧光定量内参基因的专用引物，ran基因的引物序列如下：ran正向引物5’-cgattcttgcttccctcagta-3’ran反向引物5’-accattttcgccgcctta-3’。所述的海州常山不同组织的荧光定量内参基因的专用引物，actin基因的引物序列如下：actin正向引物5’-cgattcttgcttccctcagta-3’actin反向引物5’-accattttcgccgcctta-3’。所述的海州常山不同组织的荧光定量内参基因的专用引物，hsp70基因的引物序列如下：hsp70正向引物5’-cgattcttgcttccctcagta-3’hsp70反向引物5’-accattttcgccgcctta-3’。所述的海州常山不同组织的荧光定量内参基因的专用引物，ap-2基因的引物序列如下：ap-2正向引物5’-cgattcttgcttccctcagta-3’ap-2反向引物5’-accattttcgccgcctta-3’。海州常山不同组织的荧光定量内参基因的在海州常山荧光定量中的应用。所述的rpl基因的引物序列的在海州常山荧光定量中的应用。所述的actin基因的引物序列的在海州常山荧光定量中的应用。所述的hsp70基因的引物序列的在海州常山荧光定量中的应用。所述的ap-2基因的引物序列的在海州常山荧光定量中的应用。所述的ran基因的引物序列的在海州常山荧光定量中的应用。本申请通过genorm、normfinder和bestkeeper这三个软件对17个内参进行基因稳定性评价，综合软件分析结果，筛选出海州常山不同组织稳定表达的内参基因。共获得五个稳定表达内参基因分别为rpl、actin、hsp70、ap-2和ran基因。有益效果：与现有技术相比，本发明的优点在于：本发明通过海州常山的转录组测序库，并参考了常用的内参基因，设计出了适用于海州常山不同组织的内参基因组合，同时揭露了内参基因序列，并依此设计了实时荧光定量引物，不仅解决了现有海州常山检测中没有内参基因的现状，而且所设计的实时荧光定量引物用于海州常山组织中基因表达分析时引物特异性强，从而能够大大的提高了采用实时荧光定量检测海州常山基因时的检测效率，并提高了检测结果的可信度。附图说明图1是海州常山不同组织采集的样品图；图2是琼脂糖凝胶电泳检测rna质量胶图；泳道m、1-11分别为标样、嫩叶、成熟叶、老叶、花蕾期、初花期、盛花期、果实发育初期、果实发育中期、果实发育晚期、根、茎；图3是17个候选内参基因的ct值结果图；图4是genorm确定用于准确定量分析最优的内参基因数目结果图；图5是genorm软件对17个候选内参基因表达稳定值(m)排序，稳定值越低，基因表示越稳定结果图；图6是应用cllac验证所筛选出的内参基因结果图。具体实施方式下面结合具体实施例对本发明做进一步的说明。以下实施例所使用的主要供试材料为：试验材料采用种植于南京林业大学白马基地的泰安种源、盐城种源的海州常山。在泰安种源、盐城栽培种源中分别随机选择生长良好，并且高度和长势一致的3棵植株作样品采集，每个海州常山种源样品包括根、茎、叶(嫩叶，成熟叶，老叶)、花(花蕾期、初花期、盛花期)、果(发育初期的果、发育中期的果和发育末期的果)(图1)，收集后放入离心管，收集后用液氮速冻，-80℃保存。实施例11、植物组织总rna提取及cdna的合成用easyspinplus植物rna提取试剂盒(艾德莱生物技术有限公司)提取总rna，用分光光度计(mettlertoledo，switzeriand)检测rna浓度和质量，利用1.5％的琼脂糖凝胶电泳检测总的完整性。利用北京全式金反转录试剂盒(全式金生物技术有限公司)，按说明书进行rna的纯化和合成cdna第一条链。2、内参基因的选择及引物的设计根据文献查询其他植物中主要的内参基因并最终设计出实验所需的17个内参基因，分别为actin(seqidno.1)、pp2a(seqidno.2)、rpl(seqidno.3)、pk(seqidno.4)、18s(seqidno.5)、ran(seqidno.6)、apt(seqidno.7)、sand(seqidno.8)、prof(seqidno.9)、mdh(seqidno.10)、ef-1a(seqidno.11)、ubc-e2(seqidno.12)、ap-2(seqidno.13)、hsp70(seqidno.14)、tua(seqidno.15)、ubq(seqidno.16)、h3(seqidno.17)。首先在本申请人已经获得的海州常山转录组数据库中寻找各候选基因的保守序列，根据基因保守序列用primerpremier5.0设计引物，再用oligo7进行分析。引物均在南京金斯瑞公司合成(表1)，通过普通pcr初步筛选引物，利用琼脂糖胶和凝胶成像系统观察pcr产物的条带，检测产物特异性。选择条带大小正确，条带特异性好，没有引物二聚体的引物，近一步通过rt-qpcr进一步检测引物特异性，选择峰图单一，没有杂峰，阴性对照无峰的引物作为最终的引物。表1内参基因及其对应的引物3、内参基因引物标准曲线的建立对每一对内参基因的引物制作各自的标准曲线，计算对应引物的扩增效率。将反转录的cdna混合后，以5为倍数稀释成5个梯度(1、1/5、1/25、1/125、1/625)，作为建立标准曲线的模板。同时，以ddh2o作为阴性对照模板，来检测实验过程中的试剂或者人为污染。所有的样品均做3次重复，以确保实验数据的可信度。用appliedbiosystemsstepone(thermofisherscientific，usa)进行rt-qpcr，利用得到的数据结果求得各候选基因的扩增效率及斜率，荧光定量pcr扩增效率要求所选引物的扩增效率在90％-110％之间。4、荧光实时定量使用appliedbiosystemsstepone(thermofisherscientific，usa)进行rt-qpcr，以组织样品第一链为模板进行定量分析，首先将模板稀释10倍。利用premixextaqtm(takara，japan)进行实时荧光定量，反应体系为总体积为10ul，正反向引物0.4ul、sybr5ul、cdna1ul、校正液0.2ul、超纯水3ul。扩增程序为95℃预变性30s，95℃变性5s，60℃退火30s，72℃延伸45s，40个循环，再运行溶解曲线阶段的95℃15s，60℃1min，95℃15s的分析，每个样品设3个重复，并设阴性对照。5、数据处理内参基因稳定性分析，利用软件genorm，normfinder，bestkeeper和在线分析软件reffinder(http：//150.216.56.64/referencegene.php)综合分析内参基因的表达稳定性。筛选出稳定的内参基因。6、结果1)rna提取质量及引物特异性检测各样品的od260/od280、od260/od230均符合要求，琼脂糖凝胶电泳图检测显示28s及18s条带清晰，其灰度比约为2∶1，5s条带亮度较暗，无降解现象，满足要求(图2)。17对引物在荧光实时定量中熔解曲线均只有明显的单一峰，且电泳检测也只有单一条带图，说明所用引物可特异扩增各内参基因的相应产物，无引物二聚体，且每个待测样品扩增曲线的重复性好，模板能特异性扩增，实时荧光定量结果准确可信。2)内参基因ct值分析ct值与基因的表达量呈反比，ct值越大，基因的表达量越低，反之，ct值越小，代表基因的表达量越高。17个内参基因ct平均值均在19.89-29.13之间，其中actin、hsp70表达量较高，prof表达量最低(图3)。3)软件分析genorm软件分析：genorm软件是根据平均变异度m值来衡量基因的稳定性，软件默认的取舍值为1.5，高于1.5的基因不适合作为内参使用，m值越低表示基因越稳定。此外genorm软件还根据候选内参基因的配对差异值vn/vn+1来确定合适的内参基因个数，当vn/vn+1＜1.5时，采用n个内参基因个数。根据软件，计算出v5/v6＝0.140＜1.5，所以应该在组织中选用五个内参基因(图4)，适用于海州常山组织中稳定的内参基因为rpl、hsp70、ran、actin和ap-2(图5)。normfinder软件分析：normfinder软件是根据计算候选内参基因的稳定值进行评价，稳定值越高稳定性越差，具有最小稳定值的基因为最稳定的基因。rpl的值为0.396，为最稳定的基因，h3稳定值为2.055，为稳定性最差的基因(表2)。表2normfinder软件分析结果排名123456789基因rplactinranhsp70ap-2ef-1aubc-e2sandubq稳定值0.3960.4930.6460.6530.6790.8150.8530.8671.046排名1011121314151617基因mdhpp2a18sproftuapkapth3稳定值1.0741.0891.1061.3931.5071.5411.5642.055bestkeeper软件分析：bestkeeper软件是基于内参基因ct值的标准差(sd)和调节系数标准差sd(±x-fold)来评判基因的表达稳定性，直接对基因表达ct值进行分析。sd值越小，表达越稳定，程序程序默认临界值为1，当sd值大于1时，认为该基因表达不稳定。分析结果显示基因的稳定性为ap-2＞18s＞actin＞sand＞ran＞pp2a＞hsp70＞ubc-e2＞rpl＞pk＞ef-1a＞prof＞ubq＞mdh＞apt＞tua＞h3，表达最稳定的是ap-2，最不稳定的为h3(表3)。表3bestkeeper软件分析结果4)内参基因稳定性验证综合考虑ct值和bestkeeper、genorm和normfinder3个软件的分析结果得出，rpl、actin、hsp70、ap-2和ran是适合于海州常山组织中适用的内参基因(表4)。表4综合排名选用cllac(seqidno.18)来验证所筛选的内参基因，为方便应用，只选择排名前三位的内参基因，并用表达不稳定的h3基因来做比较，发现用所筛选出的内参基因来作为内参时cllac有着相似的变化趋势，但用h3基因来做内参时，cllac基因表达情况不同，说明筛选出的内参基因是准确可靠的(图6)。序列表<110>南京林业大学<120>海州常山不同组织的荧光定量内参基因及其引物和应用<130>100<160>52<170>siposequencelisting1.0<210>1<211>1283<212>dna<213>海州常山的actin基因(clerodendrumtrichotomumthunb)<400>1ggaaagcaattgcaataaatccattccattcaaaacacaatagcagaacaacgatgtaaa60aacgttcagcgcagtccaaaatactacagcgttaaggacacgtctagaaaaccataaacg120atttttttttttagaaaagcaaatataaatagtacctaggcataccaaccaaaacaaaag180accatataaaataacttcatgcactcaaaactgcaatcgaactcacacggactccaatag240aagcatgaataaatgataccattttcgccgccttagaagcactttctgtgaacaatagca300ggacctgattcatcgtactctccttttgcaatccacatctgttggaaagtactgagggaa360gcaagaatcgaccctccaatccagacactatacttcctttcaggtggggcaaccaccttt420atcttcatactactcggagcaagagccgtaatttccttactcatacgatcagcaataccc480gggaacatagttgaaccaccactgagcacaatgttcccatacagatccttcctaatatcc540acatcacatttcattatagaattgtaagttgtttcatgaattccggcagcttccattcca600atcagtgatggctggaacaacacttccggacaacggaacctctcagcgccgatggtgatg660acctgtccatccggaagctcgtagctcttctccaccgcagagctgttcttcgccgtctct720gtttcctgctcgaaatcaagtgccacataagcgagcttctcctttacatcgcggacaatt780tcccgctcagcggttgttgtaaacatgtaacctctttcagtcagaatcttcatgaggtaa840tcggtgaggtcacgaccagcgagatccaatctcagaatagcgtggggaagggcgtaccct900tcataaatagggacagtatggctcacaccgtcaccagaatcaagcactatacctgttgta960cgaccactagcataaagagagaggacagcctggatggcaacatacatggccggcacattg1020aaggtctcgaacatgatttgagtcattttttctctgttcgctttagggttgagaggggcc1080tctgtgaggaggacagggtgctcttcaggagcgacacgcagctcattgtaaaatgtatgg1140tgccaaatcttctccatatcatcccagttgctcactattccatgctcaattgggtacttc1200aatgtgaggatacctcttttggattgggcttcatcacctacataagcatctttttggccc1260attccaaccatgacacctgtatg1283<210>2<211>1437<212>dna<213>海州常山的pp2a基因(clerodendrumtrichotomumthunb)<400>2attcccagaaactgtcatgcaagagattgtaaaaatggtgtccataaatttgtttcgtac60actctcgtcgcagcctcgtgaaaacaaagtcttggaagcttttgatgtggatgaagagga120acccttgatggatcctgcctggccccatttgcaagttgtgtatgagtttcttctaaggtt180tgtggcttcaccagagacagatgcaaagttagcaaaacgttacattgatcactcctttgt240tttgaggttattagatcttttcgactctgaagatcccagagaaagggactacctaaagac300tgtccttcaccgcatatatggaaagttcatggttcatcgtcccttcatcaggaaatcgat360caataatatattctaccacttcatatttgaaacagagaagcataatgggatcgctgaact420gttagaaattctgggtagcattataaatggttttgcattgcctttgaaagaagaacataa480acttttccttgttcgggcactcattccgcttcacaagccaaagtgcatccctatgtacca540tcaacaactatcgtattgcataacacaatttgtggagaaggactgcaaacttgctgatac600tgtcataagaggcttactcaggtattggccgatcacaaacagttcaaaggaggtaatgtt660cttgggtgagctagaggaagtgttggaagcaactcagcctccagagtttcagcgttgtat720ggttcctttgttccgccagattgctcgttgcctgagcagttcgcattttcaggttgcaga780aagggctctctttttatggaacaacgatcacatcgagaacctgatcaaacaaaaccgtag840agtgatcctaccggttatactccctgcattggagaagaactcaagaacccattggaacca900ggcggtacagagtttgaccatgaatgtccgtaaaatcttctccgatgctgatcctgagct960ctttgaggaatgtttgatcaaattccaagaagacgaagcaagggaagaagaaaccaaaaa1020gaaacgggaagcaacatggaaacgtctagaagaaatagccgcaatgaaagcttctagtaa1080tgaaccggtccttgtttctcccaggacaagcaccaagacgccctccggctaaacatggaa1140ggcttcgtttgagtcgcttgaattgtggaattatgtatttgcccctactcatgcttctgg1200ttttttttgtccggttgggtcgagcagggtgaaggaaactaatatctttttgggggttat1260gatgaagcgttgtttttgattgtaagaaatagtgggtgtacatatagaattgtgggggcc1320agatactgatgcggttttagaagtgctgatgttctatgggggaatatttgctggcaaatt1380tgccctactttcatctttagctctttagaatgatgttaaaattttaagttatatttt1437<210>3<211>1554<212>dna<213>海州常山的rpl基因(clerodendrumtrichotomumthunb)<400>3ttaaagcgaagtctcactagatctgggtcgttgtctaactgagtgataatcttatataat60taaccaaagcctgcatatagaatagggttttgagcctctggaattttactttgcggcggt120cagagtaattaggcccaagaaactatgtctcacagaaagtttgagcacccaagacacgga180tctttggggtttctaccaagaaagagggcctctcgtcacagaggaaaagtgaaggccttt240cccaaggatgatccaacaaaaccttgcaggttgactgcttttcttggatacaaagcaggg300atgacacatattgttcgtgaggttgaaaaacctgggtccaaactccacaagaaggagaca360tgtgaagctgtaactattgttgaaaccccaccaatgattgttgttggtgttgttgggtat420gtcaaaacaccacgaggcctccgttgcttgaatactgtttgggctcaacatcttagtgaa480gacctcaagaggaggttctacaaaaactggtgcaagtccaaaaagaaggctttctcaaag540tattcaaagaagttcgacagtgaagaaggaaagaaggatatccaagcccagttggagaaa600atgaaaaagtactgcactgttgttcgtgttcttgctcatacccagattcgaaagatgaaa660gggcttaagcagaagaaagcccacttgatggagatccaagtcaatggtggcgatgtagct720aagaaggttgactatgcatacagctttttcgaaaaacaggtccccattgatgctgttttt780gggaaagatgagatgatcgacattatcggagtgaccaaggggaagggttacgagggcgtg840gtgacccgttggggtgtgactcgactcccacgcaagacccaccgtgggcttcgcaaggtt900gcctgtattggcgcatggcatcctgctagggtttcattcactgttgctagggctggacaa960aatggttaccatcacagaactgagttgaacaagaagatctacaaattgggcaaggctggt1020caagagtctcactctgccatgacagagtttgataggaccgataaagatattacaccaatg1080ggagggttcccacactatggtgtggtgaaggatgactatctgttgataaagggctgctgt1140gtgggacccaagaagcgcgtcgtcacattgaggcagtctttgttgaaccagacgtctagg1200gttgcgttggaggagatcaaactcaagttcattgatacgtcttctaaattcggccatggc1260cgcttccagactatggacgagaaagctaagttttttggcaagctcaaggcttaaaatgac1320gtcgattgggtggggatataatggaatcggtttttatcgttcattttcctttataacttt1380gttggtttgccttttttcccttgtaagctgcaggtcttgaaatcgtccaaagaactttaa1440atagattttgcatttgtgttaaacctgtaccctgcacaattacaatcatctacatgttct1500catttgcgttcataagatgcatgtactggtaaatggctatacttaattaagatg1554<210>4<211>1758<212>dna<213>海州常山的pk基因(clerodendrumtrichotomumthunb)<400>4agtgataacagtaatctcctaaagtctcaatcccataccacaagtacccaccacttccaa60tctctctcctccaattacactctctcttccacacatactcacgcacttttctgagcgagg120cgcccacacattgaacacagtgagaatggcgtccacagctgcatcatcagccttctgcgg180catccgcaaggcatcgaccgccgctgccgccaaggcttctctcaatgcccctaccaccta240cttcctccccaaaacccccctccgccgcctcggattcgccgccggagccgccgaaccgat300cctatctcaccatgtcgccactaagatgcgatcatttgggaattccggcaaaccaattag360aggcgtggcctccatggcgaagaagagcatcggagaccttggtgctgctgacctcaaggg420caagaaagtctttgttagggctgacctcaatgtgcctcttgatgacaaccagaatattac480tgatgatactaggattagagctgctgtcccgacaattaagcatctgattaacaatggtgc540taaagtcattctttctagtcatttgggacgaccgaaaggcgtcacgccaaaatacagtct600ttcacctcttgttcccaggctatctgaacttctgggcattcaggttgtgaaggccgatga660ctgcattggtccagaggttgaaaaattggtcgcttcactgcctgaaggtggtgtactcct720gctcgagaatgtaaggttttataaagaggaagagaagaacgagcctgagtttgcaaagaa780acttgcctcgttggctgatctttacgttaacgatgcatttggtaccgcacacagagcaca840tgcctcgactgagggagtcacaaagttcttgaagccctctgttgctggtttcctattaca900aaaggagcttgactatcttgttggggcagtttcgaacccaaaaagaccatttgctgccat960tgttggtggttctaaggtctcctccaagatcggagtaattgaatcacttctagagaagtg1020tgatattttgctattgggaggaggaatgatatttacattttacaaagcacaaggactttc1080agtggggtcgtctctagtagaagaagacaagctaaatctagcaacatcgctccttgagaa1140ggcaaaggccaagggggtgagcctcttgctgcccactgatttggttatagctgacaagtt1200cgctcctgatgctaatagcaagattgtggcagcatctgctattccagaaggatggatggg1260attggatattggcccggattctataaagacattcagtgatgcactggggactacacaaac1320tgttatttggaatggaccgatgggagtgtttgaatttgacaagtttgctgttggcacaga1380ggcgattgcaaacaagctagcagaacttagtgggaagggagtgacaacaattattggtgg1440aggagattcagtagcggcagtggagaaagttggagttgctaatgtgatgagccacatatc1500gactggtggtggtgccagtttggagctcttggaaggcaaagaactgccaggtgttcttgc1560ccttgatgaagccacgcctgttgccgtgtagtgtgtaaaaaacaccattcttctcgatta1620ttttaacacccatctctaataatgtctcattactagcttgtaaacgagaaatttcttggt1680atacgtggtcatgctatagatgttttaccttgcctccttttcttgtcatgttaatatcac1740accatggaatggatccgt1758<210>5<211>1308<212>dna<213>海州常山的18s基因(clerodendrumtrichotomumthunb)<400>5cgaatgctgttaaatttgaatggtttaaataaaaacatattaagagagattacgcaagct60taaatctctggcaacaatcaactttcctcgaataattgtaaagggtgaagaagaaaaaaa120aatcaaaaagcaaaggccaaaatttggtatgaatagcaagtagcaacacaagcaaatttg180cttgtgtggacctaacatcttctgtgccacaatttccataattggatcgcttgcgcgact240atagagattcttattagaaccgagcttttcttttccgagcagtgtattttgaatctggtg300gcactatattcccctttctgcgcatttgttccttcttacgcagaacccattctttcccct360tcaccttcttgtttagtttttgccttttccttgctctctgccggtctgatacaaaaactg420tctgttcttcatcatcactgctatcttcatcagaactctctccatcttcacctttcgcct480ttggagtcgcactactcagagatggtgggccacaagtgagcacgaggtattctttcctcg540atttcgtactgttaccaagaagaagtattacagttaacgtgacaacttatcagaactcca600gtgcagggaagttgattcaaacctccaaggaatttacaagaataaaaccctatgcggata660atcaactactacaccaccagaaaacccagcacgcatagcaaatcccaagatcagctcacg720ttgagccaaattttcaggatatacctgtagcactgccctagctcctcgtgctaaacatct780gtataatgacccaaagaaggccttcagtctcaagcgtggctcatgagaagatttgtcagc840attgcataaccactgaacagctgagatacttattgcgccatcaataactccagggcgaag900ccctaagccctgacccatgtcaccaagtataaggtcaccctcagcttcacgctccaatgc960aatatctagcattgatcctgatatatctaaaccgatccattggtgcccatttccagccaa1020tgtctccccactaagtccagatccacaaccgatatcgagaagtaatcttggaacgccatc1080atcaggcaaagcgagaagctccaatgccctctcggaaagtttagcctgaatttcgatgat1140acgagaagaagaggtgtacttacgagcttcctcgtcgttataaaatatctcaggcggtgc1200ttgtagctctggtcgagaggccattgtttctttccttgttcttcgaaacagtactcagag1260ggtggctgaaggctgctttgttcggtgattgtccggttttgtctgatg1308<210>6<211>2008<212>dna<213>海州常山的ran基因(clerodendrumtrichotomumthunb)<400>6ccccaacatgtgggacccataaattagaatatgcatacgaagtaacagtgaattaccaaa60agtaccctcagctgatacataatgcaaaataatcaaaagtagagagatcaagggcaaaaa120tgtcacataaaatcaaacacagcacacttttggcaagcagttggcttcaggcttaaacag180cgccatctcctgctcaaatctagcagacccaccaaagacatcaaactaaaacctaatgat240ctaaatgacaaaactatcttaatctttgtcttcttcattgacatcaagattcttgagctt300cttttccaattcatcctgactaccttcgtcttcatcccccttgtcgtcaaaatcgtttcc360ttcagggtcgttctcatccaacggcccaagtttctcaggacaatctttaaagattatcct420caactcatcgagatcttcctcagaaatgaagttcccgtttacatccaatagcttgaaacc480aggcaaacgaaccaaagccagagccaacttcttaaccgaggtagttagtgagttctggct540catatcaacttccctcaattgctcttgcccatcttcaaatgctttgcttatctgaatcgc600accttcatccttaaggtcattctctgagaggttcaacttgacaatagttttctttctaga660gatgcaggccgccaagctaggagcagcttcagcagttatatcatttccagccatctccaa720tactgcaagtgatggtgctgactccttgagagcgtcggctattgctattgctccatcatc780ttcaagattcgagtagcttaaataaatctcagttagattcccatgtttggagagagcgtc840gcttagcttcaaaccagcctccactccaaacatattgtcccgaagatcgattttcttcag900attcttacataattccagggcttcagtcaacgcaacgcctccttcagaaccaatacgggt960agatgagcatcggaaatcctccaataaagggcaacgtttcagtatctcggaaatggcaat1020agcgccttcatctcctgtcatattgttatgaaaatggaggattctaagcgaatctgtaga1080aggaaccaactcacaaacagcttgtgcggcttccttcgatatcccatcattcatcagata1140aagttcctctaagctagtctgagatttcaagagcttcccgaatgctctgactcccttctc1200acctaaggcattatcggagatattgagatatttcaaatgagaaccttctagagcttcaga1260gaagatcttcatgacatcaagagcttctgcttctggtcgccccgcaacaaaatcagacag1320atcgacttccttcagttggttcttgattgacgccaagataggtccagcaacattggctgc1380cccaagaccaaagcttcgattgctaaagcaaatcttggtgtaagaattcccaggttcttt1440caaggggcttaaaagtgtttgagcctcatctgcttcaataaatgctcgctgacccttgga1500aatatcgaaaaatgtctcctgcaaaaccgaattgacctcagatttcagtacctgcttttc1560ctctgtcccagagcctcccttgagaacatccagtattagcttgctgcattccttggcata1620ccattgtacagcagaactgccatcaccatcgggttcttgttcataatgctgatttgccga1680agaaaatgctgattcttcaatttgtttggcatgttttgaagcctcagcttgactaagagt1740gccatacttgcgggtgaaaactgttggcgtcgagagattgtttgtcattcgctctacaag1800catcaacctcgtgttctggctagggggccacagtttaatggttatgtgccggcgttctcc1860atttggtgttttagcatccatcgcacccaaactgtcgtctccttcctgacgaatttttca1920atttgcagctcttcggacttcgactgtgtactgcctgtgtatgtataggcaagaggggag1980aagtgaaggaaaactaaaatgaaatgag2008<210>7<211>1149<212>dna<213>海州常山的apt基因(clerodendrumtrichotomumthunb)<400>7aaaatctagagatttaggcaaatccattagaaaacaagcatcctgtcttcaattgattct60aaaaatcccactaatgtacgctgtatctgtatctaagccatctctctccctatcaagtga120tgtcattgattaaacaaatcttgagttttaggcaaatccattagaaaacaagcaccctca180tggacttcaaaaatcccactaatgtatgctgcatctgtatctaagctatccctcaaactc240ctcaaggctagagttgtctcaccaaagtacttgcgcagtgttaatttctgcttacctcac300accgtagcgaacactccgctgacctacgtcggatcaattggtggcgcagccgtaccgaaa360cgtcggccggtgttcgtttcaaacatggcatcctcaaatgctgaaagcattgatgagcgc420atccggcggattgcctccacgattcgggtcatacccgactttccaaaacctgggatcatg480tttcaagatataacgacgttgttgcttgatcctaaggcgtttaaggatacgattgatttg540tttgttgagagatataaatgccaaaacatcaatgttattgctggtgttgaggcaaggggt600tttatattcggtcctcctattgcattggctattggtgcaaaatttgttcctatgaggaag660cccaaaaagttacctggggaagtaatatcagaagagtactctttggagtatggaactgac720ataatggagatgcatgttggagccgtacaacctggggaacgtgcagttgttgtagatgat780cttatcgcaacagggggtaccttgaatgctgctattcgcctacttgaacgcgttggagtc840catgtggttgagtgtgcttgtgttattgaactggagggactgaagggtcgggacaagtta900ggggagaatccactgtttgttcttgtgagctcaccttgaagaaaagagtatctacaacaa960caataaattgctgttaacatctgtatgagttgatagcatgtgcttgttcctttctatggg1020atatacacatttcaaatggatgatggtgtgttgtatgttgagtgaaattgcttttctttt1080ttttttggatgagtcatgttcaggccaccctctcctgcttgcggattgttactcaacctg1140tcaaataag1149<210>8<211>2473<212>dna<213>海州常山的sand基因(clerodendrumtrichotomumthunb)<400>8gtaatttgtttctgtctccgccatccgtgactggcagatgcgagtgcgacaccatcactg60aacttcaaatgttttcctttaaactctaatttcttagcgattatccaatccaattctcgt120tcgggattgtagattgattgacaatgtcttccaattcaaatatttcatctgcgagttcct180ctactaatggagatgacgggaaccctaattcgtatacaaaaaaggagccggagaatctgc240cggaacccgcactcaacacgagtccggacttgatcgaccagtcgcttgatgcccttgaga300atcaattggcctcaatatcaatgtcctcccgttccccgacggcgactcttgacgatgaag360agttcgtcgaagaagaagagcagtacctggagcatccaacaatcataaatggatcgttga420gtgaagaaaatggagccggaggatcaagtgatgagcatattgagtgttatttgaagggag480agtcttcgggagtgataaaagagaccttgtcctctgtgtggaggaattatttagaggcag540atgaagttgagcggccgttgagtccgaccagcagcggctacgctggtgagatggggagca600ccagcggcggtgatgaaatcaatgatgatgacgaagatattaatgaaattaaagaggtcg660aaaacgactttgtttcggggtcccatgatcagtgggtttcaggaaaacgccacccagatg720aggatgatgcttccgtttcttggaggaaaaggaagaaacatttctttatcctgagtcact780ctgggaaacccatatattcgagatatggagatgaacacaagctagctggattctctgcaa840ctttgcaagccattatttcctttgttgaaaatgggggagaccgggtcaaattggttagag900ctggtaaacaccaggtgatttttcttgtgaaaggacccatatacttggtttgcataagtt960gcacggaagaaccttatgaatcactaaaagaccagatggagctactctacggtcagatga1020ttctcatcttgacaaagtctgtaaatcgttgttttgagaagaatccaaagtttgatatga1080catcattgctggggggaactgatgctgtcttctcttctctcatccattctttcagttgga1140atcctgccacttttctacatgcctactcatgtcttccccttgcttatccaacacgtcaag1200ccgcaggtgccatcttgcaagatgtagctggttcgggagtcctatttgcgcttttgatgt1260gtaaacacaaggttatcagtcttgttggcgcccaaaaagcatctcttcatcctgatgata1320tattattactgtcaaatttgataacgtcctctgaatcttttaggacatctgaatccttct1380caccggtttgcctgccaagatacaatcccatggcatttctttatgcttatgtgcattact1440ttgatattgatacatacctgatcttgctcaccacaagttccgatgccttctatcatttaa1500aagattgcaggattcgaattgaaaacgtacttttgcagtcaaatgtactgagtgaagttc1560aaagatccttggtggatggcggtatgcatatagaggatttgcttgctgaccccacatctc1620gtcctggatccgtttcttctcatttaggtcaaccagggcctggtacaggttccccggggc1680gacttacagatggatttgttggaattggtggtccagctgggctatggcatttcatataca1740gaagtatttatcttgaccagtatgtgtcatctgaatttttatcaccaatcagcagcttga1800gacagcaaaaaagattgtatagagcgtaccaaggactttatgcatccatgcatgacaaag1860gaattggacctcacaaaactcagtttagaagagatgaaaattatggtgacatttactaaa1920gcaaccaaattcttcaattatatgcaagactggaactctggaagatagtttacatcgaga1980ttctatggggtagagccataaattgaagccatgctactctgctgggtcacacaagatttt2040gaactatatgcagcatttgatccactagcagacaaggcacttgctataaaggtgtgcaac2100cgggtttgtcaatgggtgagagacgttgaaaatgaaatatttttattgggagtaagccct2160ttttcgtggtgattatctcaagtattctgtacactggcatgtattctagtctttccgttc2220gactattacttaactaccgcttcctatatgaaattttgccttatttttgttgtacggcct2280ctatgttggtttgcagttcggatctctatcctattttttcttcaccacttgtatctaact2340ataaaaacgaataaattgaagttctatagtgagcttaattgtagttgttgtaattggatg2400tttataaaaaggtaacgttttaacagtgagacataagtggtatatggggaagtatgattt2460tagaggataccga2473<210>9<211>1456<212>dna<213>海州常山的prof基因(clerodendrumtrichotomumthunb)<400>9actaacttgataccaatacgcttgcgattaattgttatctccgccgagagagaagctctc60gcatctttctaagaaagaaattttcactttagatctatttgttttcactctccatcctca120cctccctatctttccgacgttgaagtctcttcctcacttcttgatcctatactaactcca180gtttctacatttgaaatgaagatggaagatctttttaggtctagacgcaaccagagttgt240gtgacagcagcagcggcaagattagggcgactgcagaaggtggatgtagttccaccatgg300cagtgacccttgatgtgtgtatttccaaattttctttagtgtagttttacgttgttttca360tgttcatcctcgatctggactggttctcaaggtgctagttttagggtgctttatactagc420tagtcaagttgggacagtgacctgaatctctctctaccaatgtccttgaatagggttttg480gtccggtcaagctttctttattttgtctcctttagtacgagtggtttttcttccatcttt540gggattgaccgttcattttgtacatttagtcagttggctatgtattccacatgtatgtga600agttttgggggaagaagataactaaggggaaaaaatttcaaagccaattctgtatttgta660aagatgtccataaaatttgaacatctatcagagaaccaaccaaaaatacataatttactc720acatggaatcataaacccaagttttcacaacgagaaaatcaaaagacaagccctgtttca780attcagatagggacataatctaacacctctcactcaaaacaccacattctttttgccata840accaacaacccacgctgcaaatcattgacaaaaaaagcagtaaaggatgccgaatttaat900aatattacccaaaaaattattaatttataattatataatgaatattgattattaaatgaa960atgatttggaagacttaccaatcaatttcagacaccctgttcaataagatagtcaccaat1020cttctcaacaaccatgttgcactggcctgcagtcattggttcatcatagataccgatgat1080caaggccaaattggtctttttgacagtggcaccaccagatcccttcttccctcggatgac1140agcatttggttcaccttgaatcaccatgtatttagtacccccaatgtacaatccagtagg1200agctaatgatcccggattatcaaaatcattcataatagctgttacttcctcaggcttgaa1260ctgagggaagttggaactctgtgcccaaacgctgccgtcatgaccgacaattgcggcgga1320ggtgagatggcaaccgtcgacgtcggccatcaagtggtcatctatgtaggattgccacga1380catcgtttcgataggcggaaggaatcaataccaggaaggtttggagaatgggtgaggact1440gatgatagcgtgaaaa1456<210>10<211>1516<212>dna<213>海州常山的mdh基因(clerodendrumtrichotomumthunb)<400>10tttataagaattggtaacatccattggtatatgatatttatctgtcagcaaacgggttca60ttacatatctagaattgcaataaccagaacaactcacgtagcataaggtacaacatacac120taccgcttattccaagatgtgtgatcttgaatctccaagattatttccttacgaaggaga180cccccttatcaatgcttccatccaactctttctttgccttctccagcccaaccctttcgt240attcatttagtggaccaagagggaagatctcctcgaccccgttacgtccaagccgtaccc300ttgaagcaaagaatggaagttccgttacctgggaagaaacaaatgcacattcaacaacac360cggaatctcccctcaagcccttcaggcaagcatcggcaaatttcactgctgcatatgcca420tggaaagagttgcagagcctgtacccgctttggcctcaacaacttcagtcccaccattct480ggatgcgagatgtcagatactcagtttcttctttggtgaaagagcatgcaggtgtaatct540gcgagaggagaggtaaaattgtcacaccagcatggcctcctacaactggaacattaactt600cccgaggatctagccccaaaacttcagccacaaacgtattagctctgactacatcaagca660tcgtgactccaaggagtcgccttggatcgtaggtcccagccttcttgaaaacctctgcag720caattggaactgtggagttcactggattacttattaagttaacaatggccttagggcagc780acttagcaactccttcacaaagtgtctttactattcctgcattaatgttgaaaagatcat840ccctcgtcattccaggtttcctgggaacaccagcaggaatgattacgaggtccatgccag900ttagtgcatccttcaactggttttgacccagaaaaccacgtaccacagcaccagtgttca960tatggctgatatctgctgtaaccccaggagtattaactacatcataaagatgaagcaccg1020aaaccagtgggttcatcttcatcagcattgctaaaggctggcctatgccgccagctgccc1080ccaataccgccaccttgaaacctggtgaccctccttttgctcggcaatcggaaagccata1140aactcccctccacctgcagctgattagtaggaggatcgaggtgagctgaaattctggaaa1200ttcttcgatgaatatccccaccctgctgcatactataactcgaacgaattcttctccaat1260atcagaaagcagcaaattccttttggaataatgagaaatgaaatgatggaagataacttg1320aacggcagaatagtggagatacccgtatttatacggtagaaaattggaattaattgtatt1380ttatggcacagcacataataaaataaataaatacgtcagaggctcatagaaatgcgttaa1440tgtatttttgggtgcatagccttatcttatacgtacacgcttcggcaagaaaattacaaa1500ttatcaccataattca1516<210>11<211>1351<212>dna<213>海州常山的ef-1a基因(clerodendrumtrichotomumthunb)<400>11gaaattcgagaccaccaaatattactgcacagtcattgatgcacctgggcatcgtgactt60catcaagaacatgattactggaacctcacaggctgactgtgccgtcctcatcattgactc120cacaacaggtggttttgaagctggtatttccaaggatggacagacccgtgagcatgctct180gctggctttcactcttggtgtcaagcaaatgatctgttgttgtaacaagatggatgccac240tacaccaaaatactccaaagcaaggtatgatgaaattgtgaaggaagtgtcctcttacct300caagaaggttggatacaacccggacaagataccatttgtccccatttctggttttgaggg360agataacatgattgagaggtctaccaacctagattggtacaagggccccacgctactcga420tgcacttgaccagattaacgagcccaagaggccaacagacaagccactccgtctcccact480tcaggacgtttacaagattggtggcattggtactgttccagtaggccgtgtggagactgg540tgttctaaagcctggtatggtggtcacctttggcccgtctggtctcaccactgaagttaa600atcagttgagatgcaccacgaagccttgcaggaggctcttccgggtgacaatgtcgggtt660caacgtcaagaatgttgctgtcaaggatctaaagcgtggctttgtggcctccaactcaaa720ggacgaccctgctaaggaagctgcgaacttcacctctcaggtcatcatcatgaaccaccc780tggccagatcggaagtggctatgcacctgttctcgactgccacacctcccacatcgctgt840taagtttgctgagttagtcaccaagattgataggcgttcgggtaaggagattgagaagga900gccaaaattcttgaaaaatggtgatgctggattcgtgaagatggttccgactaagcctat960ggttgtcgagactttctctgagtacccacctctgggtcgttttgctgtacgagacatgcg1020acagactgttgctgttggcgttattaagagcgtggagaagaaggacccgagtggtgctaa1080ggtgaccaaagctgctgctaagaagggtgctaagtgaacgctggtgactccgttttatca1140ataaattttgtttttataatgcagtttatgattaatgattattgcttttactgctgtatc1200tctagcctcttgtaagaggatattggagagtggagcaacttggttttgattcttgttgct1260tcatgtgcagtgatgaaagcagctctgtttggatattgtcaaatttttattttccttgtg1320gattttgttttcttgaaattgttacttagtg1351<210>12<211>1214<212>dna<213>海州常山的ubc-e2基因(clerodendrumtrichotomumthunb)<400>12aaattgttttgcaaaaacctcgattttcatagtactcgttcgtcggtgtgcgatctgagg60agatggcggaagataagtataacaggaagaaccctgctgtgaagaggattttgcaggagg120tgaaagagatgcaatccaatccttccgatgatttcatgagccttcctctcgaggagaaca180tatttgaatggcagttcgcaataagagggccaagagactcggagttcgagggaggaatat240atcatggaagaattcagttgcctgctgaatacccctttaagccaccttcatttatgttat300tgacgccaaatggtagatttgaaacccaaaccaagatatgcttgagcatttcaaatcatc360accctgaacactggcaaccatcctggagtgttcgaactgcgctggttgcattgattgcat420ttatgcctactagcccaaatggtgcgctaggatctttggattacacaaaagaagagaggc480gagctcttgcagtaaaatctcgcgaagcagctccaaaatttggatcacctgaaaggcaaa540ggctgattgatgagattcatgagtatatgctcagcaaagcgctgcctgtgcctcaaggca600acctcacctctgaagcacacactgggagtgaaaaaatccagcagaccacccaagacaatg660ttgaggaagttacagaagcacctccccacacagctgaagacgacgggatcatacaagaac720caagtgaagcagtagcaccttctactaatacaaatgttgctcagacatcgcagcctgttc780ctcccgggccgacgacgagccagcagcagctgttgcataagcccgaaccaagggttccta840aaccagcagatgatcgtctcttcacatgggctgctttcgggctcaccatagctatagtag900ttcttctgttgaagaagttcttgaaggctaatgggcacggtgctgtctttatgaatgagt960cgtaagtgggacctggtatgttgtttatgctgatgcagaaggtgaatgaggggaaaacct1020gctttctgtaatggagtgccttaaattttaaggggaacatatctatatatatattatacg1080tatacttatctttctacttatggagtatattgtatagtttaaataattgttaggggtaat1140gttctgaagctttttaagaagctgaactattttaatagaagcagttctgtaattattgct1200tgcatttgtgagag1214<210>13<211>2673<212>dna<213>海州常山的ap-2基因(clerodendrumtrichotomumthunb)<400>13ttaatcaccgatgtcttcagaaccaatacgcaactaattgacaaacagatgctgtaattg60ttacaaagggtgcagttttcattatgtcacttgcattacagaaaccaatgaagaagggtg120tatttggcaagcgctaaataaagaggaaaaaaaatctcaccaaggcacaatctgattgtg180cgaaagaattatcaaaacttcaaatccaaattctaacactatgccgattacatacaaaat240tacaaattcaccctccgactcccccagctcttgcaccatattagcggggatccctccagt300acatacgagcaggtgctatatgaacattggttaaacaattttgaacagccttgggtttgt360tttatcactcaaagcaaaccagcaagaatagctccaggatctgatgtagatgtaggtgga420ggcgttagttgtggttgtggttgaggttgaggttgaggttgaggttgaggtggcgtaggt480gcaggtgctcgagaagggatgggaatggtgattaattgttctttaatgaattctttcaat540tcgtatgttagttttggatctccagatgcgatggtcatccgtaattgcgttctatctgct600gggtctgtctcgattcttatcaaacataacatggctcgtgtactttcagagtagaaagtt660gtgctcgcaaccaaattattagcatttggatcaagcccagggcaaaccatcaggtgcaaa720ctgctaaataagtttgccatctctgctagcagcattggtcttacacctctaaccacttct780tgcagtttcaatggaggtcctgatagtgatctccattgtgggaaaaattcttcagcagaa840actgtaattgactgaagaaatttgttcaaaacagcaggaaggcgaagttttacattgacc900aaatgggttccaaacttgtacgagaagtccagtactgctaaatctctgctcggttggagg960atgattacttcaagtgggcattgaacctgtgcccttggaggaatagtctcaggtactaaa1020gatagttccatctttagatgtgatggaggcaatattaaagcttgaactgaaacgagtgga1080gcagtatttttatttcccaagaagagaacaactcgcccatgatgggttcgccattcagct1140tttatgccaatctggatataggcatcttcatatagcacaccactatccttcaggcacaaa1200acacgaaatctttccactatatcaccaatcggctggaccgtattaacttgctcttcaaca1260ggtgctatggctaatgcatctgtttccaaaccggaagcaacccttggatcagtttgggga1320gcaatgcctggtgggccctcaatagccagtggccctaaaagatctccaaggagatcagga1380gaaggtgcactgtgtggttgaggatccactacactcagggcaccattagcacgagtcaac1440ccttgctcagcagtgctctgatccgcattactcatactaggcaccttgacaagaccaagc1500tgggtcccagatggtgttccatttgcagggcgttgatgatcggtaagtaccaaagcatta1560gaattctgctgctgtgcccgtaattttattgcactttgctcagcagtatcagcttcagca1620tcttccgctttttttatcaatgatgactgtctctcagggaatttcggcatttcagcaagt1680atatccactagagcagcacctttcatgctcagagcaagatactcaacagctcgttgctgt1740atttcagcatcaatgcaactttcatactttttgaatatcacccatatttgcttctgtagc1800tcctggctgggtggttgagtgtgcatcaaaatttttgcatatgtggaaagaaggatagga1860attgttgatgtcgagacagtgggaagcttctcatgtatcaagttgaaaatttcctttgga1920ctgcacccaggtcgcctagctagaagatgactgtattctccaaggatatatgcactcacc1980ttcaccatggtctcgtgaatcgcaggcttatcaagatactctttggctttcagagctgca2040taaggctgtaaatcctcgttgtttgtaacgaactgcacgactcgaaaccaaatgtcatca2100ccgacaaaatcccctgccttgtcaattagctgaagaatcacatccacataccatgacagg2160tcgggggcaaacttctctgcgagaatagcagctttaagtgacaattcttcccgcatagca2220aagtcagctgagctcagatactgtaataattcttcaactatgtcctttgcatttgaaaca2280tcacacattccatatagcaaatcaagggcacgccttctgatactgatatcaggatccttc2340agggaagtaatgatctgagcttgatgtcttttgataatttcatgcacatcggcgaccatc2400aacattcgagtcatgttctccaagccaaggtatcgtatattaggttcacgaacagcaata2460aattttcccagcaaagcaacacactgagacatcatttctttctctgcatcaagatgcatg2520acaagagcaagagcttcaaaaagaacagcatgtgatgcattattcttgttcacatttttc2580accacatcagttcccatgagtatccgttgcagaacctcaaacaatgatcttcttgtattt2640ggatcctcaactgttggaaaatactggagagcc2673<210>14<211>2071<212>dna<213>海州常山的hsp70基因(clerodendrumtrichotomumthunb)<400>14tgttagttttatttcattattgttctttgacttgagcaaaattctctggtcgatttagat60ctgcgattagttttgcgagcaatggccggaaaaggagaaggtccagcgatcgggattgat120ctcggaacgacgtactcttgtgttggcgtttggcaacacgaccgtgttgaaatcatcgcg180aatgaccagggtaacagaacgacgccgtcttatgttggcttcaccgattcagagaggctt240attggtgatgccgctaaaaatcaggtcgccatgaacccgacgaacactgtttttgatgcc300aagaggttgattggaaggcgatttagcgatgcttctgttcaaagcgacatcaaactatgg360cccttcaaggtcattgccggacctggtgagaaacccatgattgttgtcaactataagggt420gaagacaagcagtttgcagctgaggagatttcatctatggtcctgatgaaaatgaaggaa480atagcagaggcctaccttggttccacaataaagaatgctgtcgtcaccgttcctgcatac540ttcaacgactcccagcgtcaagccaccaaagacgctggcgtaatttcaggtctcaatgtt600atgcgtatcatcaacgaacctacagctgccgccattgcttatggtcttgacaagaaggcc660tcgagtgtaggtgagaagaatgtcttgatttttgatcttggtgggggcacttttgacgtc720tctctgttgacgatcgaggagggtattttcgaagtgaaggccactgctggcgacacccat780cttggaggtgaggactttgacaacagaatggtgaaccactttgttcaggaattcaagagg840aagagcaagaaggatatcaccggtaacgctagagctctacgaaggctgagaactgcttgt900gagagagcaaagagaaccctttcatccacagcccaaacaacaatcgagatagactctcta960tacgaaggaatcgatttctactccaccatcacccgtgcaaggttcgaggagcttaacatg1020gatctgttccgcaagtgtatggaaccagttgagaagtgcctgcgcgatgccaagatggac1080aagagcacggtgcatgacgttgtgctcgtcggtgggtccacccgtatccccaaggtgcaa1140cagctgcttcaggacttcttcaacgggaaggagctttgcaagagcatcaacccggatgaa1200gccgtggcctacggcgccgccgtccaagccgccattttgagtggtgagggtaacgagaag1260gttcaagacctattgcttctcgatgtcactcccctctcccttggtttggaaactgccgga1320ggtgtaatgactgttttgattcccaggaacaccaccattcctactaagaaagagcaggtg1380ttctctacctactcggacaaccaacccggcgtcttgatccaggtctacgagggtgagcgt1440acgaggacgagggacaacaacctgctcgggaagtttgagctctccggcattccccccgcc1500cctcgtggggtcccgcagatcactgtgtgcttcgacatcgatgctaacggcattctaaac1560gtgtcggccgaggacaagacgacgggccaaaagagcaagatcacaattacaaacgacaag1620gggaggctgtcaaaagacgagatagagaagatggtacaggaggcggagaagtacaagtcg1680gaggacgaggagcataagaagaaggtggaagcgaagaatgcgctggagaactatgcgtac1740aacatgaggaacacggttaaggatgaaaagatcgggtcgaagctgcccggtgctgataag1800aagaagattgaggatgcggttgagtcgacgattcaatggctcgatgggaaccagctcgcg1860gagtcggatgagtttgaggataagatgaaggagttggagggcatttgtaacccgattatt1920gccaagatgtatcagggtgcggggggtgcggcggctatggatgacgatgacggtctggcg1980gcggcgcctggtggtgctggtcctaagattgaagaggttgactaagttgtggcccctttt2040ttggtggtttttttttttttttttttttttg2071<210>15<211>1802<212>dna<213>海州常山的tua基因(clerodendrumtrichotomumthunb)<400>15atctgagccctagagagagaaagtgaaattacaccttcattttctaccaactgctcttat60aaaatcccctttcaccacgaattaagaactacctattttcatccagagacgtcttcgtat120acgccctggtatttctctcctttgatctccgatcgaaatccatttcagattttaatcttt180caaacaaaaagcgaaaatgagagagtgcatttcgatccacattggacaagccggaatcca240ggtcggcaatgcctgttgggagctgtactgccttgaacatggcatccagcctgatggaca300aatgcctggtgatcacactgtcggaggaggtgatgatgccttcaacacattcttcagcga360aactggcgccgggaagcatgttccccgtgcagtgttcgtcgatctggagcctactgtgat420tgatgaggtgcgcactggtacataccgtcagctattccaccctgaacaactaatcagcgg480caaagaagacgctgccaacaactttgccagaggtcattacactattggcaaagaaatcgt540ggatctgtgccttgataggatccgcaaactcgctgacaactgtactgggttgcaaggctt600cctcgttttccatgctgtgggtggtggtaccggatctggcctcggatctctgttgctcga660gaggctctccgtcgattatggtaaaaagtcgaagctgggtttcactatctacccttcccc720tcaagtctcaactgccgttgttgagccttataactctgtgctttcaactcattcccttct780tgaacacactgatgttgctgttcttcttgataatgaggccatctatgatatttgccgcaa840atctctggacatcgagaggcctacatacaccaatttgaacaggctcatttctcaggtgat900ttcctccttaactgcatccctgagatttgacggagcattgaacgtcgatgtaaacgaatt960ccaaaccaacctcgtcccatacccaagaatccacttcatgctttcatcatacgcccccgt1020aatctcagctgagaaagcctaccacgagcagctctccgtcgctgagatcaccaacaccgc1080ctttgagccatcttccatgatggtgaagtgcgatccgcgccacggcaagtacatggcgtg1140ctgcctcatgtacagaggcgatgttgtccccaaagatgtgaacgcggccgttgccacaat1200caagacaaagaggacgatccagtttgtcgactggtgcccaactggcttcaagtgtggcat1260caactaccagccgccgactgtggtccctggtggtgatcttgctaaggtgcagagggcagt1320ttgcatgatttcaaactcgactagtgttgctgaggtgttctcgaggatcgatcataagtt1380tgatttgatgtatgctaagagggctttcgtgcattggtatgtgggtgagggtatggagga1440aggggagttctcggaggcgagggaggatttggctgctttggagaaggattatgaggaagt1500tggggctgaatctgctgaaggtgaagatgatgagaatgaagagtactagagtagggacca1560attggctcgttggtttagtgctgaacttttttccttatggttgaacttcttgtctggtgt1620tgaactgcttatggattctctgtgtgtagttcgtattgattatggtatgtgttgaaaagg1680ggctttatgttcatttcgtattgattatgcttgtgtaattcgtatgtgttgattattagt1740tattccctaggtcatactcgtattttatatgtttaaaatgccaagataattttttttttt1800tt1802<210>16<211>1670<212>dna<213>海州常山的ubq基因(clerodendrumtrichotomumthunb)<400>16gaattgttctcataaattttcagttttttcgtctctcaatcgtcaagatgcaaatcttcg60tgaaaactctcaccggcaagaccatcacccttgaggttgaaagctccgataccattgaca120acgtcaaggctaagatccaggacaaggaaggtatccccccagaccagcaaaggctgatct180tcgcaggaaaacagcttgaggatggccgtaccctcgccgactacaatattcagaaggaat240caactctccacctggtcctccgtctccgtggaggtatgcagatattcgtgaagaccctaa300cgggcaagaccatcaccctcgaggttgaaagctccgacacgatcgataacgtgaaggcca360agatccaggacaaggagggtattcccccagaccagcagaggctgatcttcgccggcaagc420agctcgaggacggacgtaccctcgccgactacaatatccagaaggagtccaccctccatt480tggtcctccgtctccgtggtggtatgcagatctttgtcaagaccctcaccggcaagacca540tcaccctcgaggttgaaagctccgacacgatcgataacgtgaaggccaagatccaggaca600aggagggtattcccccagaccagcagaggctgatcttcgctgggaagcaactcgaggatg660gacgcacgcttgctgattacaacatacaaaaggaaagcacacttcatttggttctgaggt720tgagaggtggaatgcaaatctttgtcaagactctcacggggaagaccatcaccttggagg780ttgagagttccgacaccattgataatgtgaaagccaagatacaagacaaggaaggtatcc840caccagaccagcagaggctcatatttgcagggaagcagctggaggatggacgcaccttgg900ctgattacaacattcagaaggagtctactcttcatttggtgttgaggttgaggggaggga960tgcagatatttgtcaagacattgactgggaagaccatcactttggaggtggagagttcag1020acactattgataatgtgaaggccaagatccaggataaggagggcatcccacccgaccagc1080agaggttgatcttcgccgggaaacagctggaggatgggaggactctcgctgattacaata1140tccagaaggaatccacccttcaccttgtcctcaggctccgtggtggtttctagagcttct1200ggttttcttgttgtttgaatgttctggttttatgttctgtttgttgattttacatgattc1260ctgttttcccgtgtgtgtgtgcgtgtgttgaaacttaagtatggtactgttatgttgtcg1320tcttgctgacattctcagttgtcattggaataaaattgggattttatcccccctataata1380aatttccggtgagtgcataattggctgtaatagttttttcaatattgttgctgttgagaa1440ggtgctacgtaatgttctaccttcttcatcggttgcttttctggagattcttgtatgcct1500cggtaatgtttggtgattatatttatcttctttaattactgaaagaagttggatccgatg1560cactatcccaggagaggtcgaagcgatgtattcctaggtgtttttctgttagtggagttt1620acttcgttgcatgttgtcccgtcaaccattttcgtaagggattaatgttt1670<210>17<211>588<212>dna<213>海州常山的h3基因(clerodendrumtrichotomumthunb)<400>17actgatcagttttactacaaaacagacaaaggttaaacattcaaagccacaagtgacacc60tcgaaacaaccacatatttaccatcggaaaaaactatcccttagacccaaatcatccaga120ttacaagttaagccctttcacccctaattctcctcgcaagctggatatctttgggcataa180ttgtaaccctcttagcatgaatggcacagaggttagtatcctcaaacaatccaacaagat240aggcctccgctgcctcttgaagagccgccacagcagagctctggaacctcagatccgtct300tgaaatcctgagcaatctcacggacaagcctctggaaaggaagttttcggatcaaaagct360cagtggatttctggtacttgcgaatttcacggagagcgacggttcccgggcgaaaacggt420gaggcttcttcactccgccggtggcgggggctgatttcctagcggctttggtggccaact480gcttccgcggtgccttgcctccggtggatttgcgggcggtttgcttagtgcgagccattt540ctagggtatcgatagaaagtatggaaaaaaagtgctaatttggaattg588<210>18<211>514<212>dna<213>海州常山的cllac基因(clerodendrumtrichotomumthunb)<400>18gccaataatttctcagcaatggagggcttattttaccgtcttctccttcttacagttcca60ttaatggccatgcaagtgcttccttcttcttcatcatccaatacaaagatgtttcagttt120aatgttgaatggaagagtataacaagactttgcaatagaaagacagtattgacagtaaat180gggaagtacccagggccaaccattgctgtgaatgaaggtgacagtgttcaagtaacggtc240actaattgggttgcaagaaacaccaccatccattggcatggagtaaggcagatgagaagg300ggttgggcagatggggcagcatacataacacaatgcccgattggaggaggcaaaagctat360agatacaactttagtgtagaggatcagaggggtacactgtggtggcatgctcatatcggc420tggcaacgtgcttcggtttatggcgcttttatcatttaccctcgcacgccttttccattc480tctctccaactccacccacttcagattcctctca514<210>19<211>21<212>dna<213>actin引物序列(artificial)<400>19cgattcttgcttccctcagta21<210>20<211>18<212>dna<213>actin下游引物序列(artificial)<400>20accattttcgccgcctta18<210>21<211>19<212>dna<213>pp2a上游引物序列(artificial)<400>21ggacaagcaccaagacgcc19<210>22<211>22<212>dna<213>pp2a下游引物序列(artificial)<400>22tcagcacttctaaaaccgcatc22<210>23<211>21<212>dna<213>rpl上游引物序列(artificial)<400>23agtcaatggtggcgatgtagc21<210>24<211>22<212>dna<213>rpl下游引物序列(artificial)<400>24cccttggtcactccgataatgt22<210>25<211>23<212>dna<213>pk上游引物序列(artificial)<400>25ctgaaggtggtgtactcctgctc23<210>26<211>21<212>dna<213>pk下游引物序列(artificial)<400>26cgaaactgccccaacaagata21<210>27<211>21<212>dna<213>18s上游引物序列(artificial)<400>27acaatcaccgaacaaagcagc21<210>28<211>22<212>dna<213>18s下游引物序列(artificial)<400>28cgagaagtaatcttggaacgcc22<210>29<211>20<212>dna<213>ran上游引物序列(artificial)<400>29ggagacgacagtttgggtgc20<210>30<211>19<212>dna<213>ran下游引物序列(artificial)<400>30tgccatacttgcgggtgaa19<210>31<211>19<212>dna<213>apt上游引物序列(artificial)<400>31ccacgcattcaagccattc19<210>32<211>20<212>dna<213>apt下游引物序列(artificial)<400>32gagcctctatcccagcaacg20<210>33<211>19<212>dna<213>sand上游引物序列(artificial)<400>33gaccagtcgcttgatgccc19<210>34<211>22<212>dna<213>sand下游引物序列(artificial)<400>34ggatgctccaggtactgctctt22<210>35<211>24<212>dna<213>prof上游引物序列(artificial)<400>35gaatgtggtgttttgagtgagagg24<210>36<211>18<212>dna<213>prof下游引物序列(artificial)<400>36atgtgaagttttggggga18<210>37<211>20<212>dna<213>mdh上游引物序列(artificial)<400>37acctggaatgacgagggatg20<210>38<211>24<212>dna<213>mdh下游引物序列(artificial)<400>38gccacaaacgtattagctctgact24<210>39<211>20<212>dna<213>ef-1a上游引物序列(artificial)<400>39aggatggacagacccgtgag20<210>40<211>22<212>dna<213>ef-1a下游引物序列(artificial)<400>40aaaaccagaaatggggacaaat22<210>41<211>23<212>dna<213>ubc-e2上游引物序列(artificial)<400>41gcaaaggctgattgatgagattc23<210>42<211>21<212>dna<213>ubc-e2下游引物序列(artificial)<400>42cctcaacattgtcttgggtgg21<210>43<211>20<212>dna<213>ap-2上游引物序列(artificial)<400>43ccacaactaacgcctccacc20<210>44<211>20<212>dna<213>ap-2下游引物序列(artificial)<400>44aaattcaccctccgactccc20<210>45<211>20<212>dna<213>hsp70上游引物序列(artificial)<400>45gccattttgagtggtgaggg20<210>46<211>23<212>dna<213>hsp70下游引物序列(artificial)<400>46ggttggttgtccgagtaggtaga23<210>47<211>20<212>dna<213>tua上游引物序列(artificial)<400>47gtccccaaagatgtgaacgc20<210>48<211>20<212>dna<213>tua下游引物序列(artificial)<400>48aagcagccaaatcctccctc20<210>49<211>21<212>dna<213>ubq上游引物序列(artificial)<400>49tgctgttgagaaggtgctacg21<210>50<211>19<212>dna<213>ubq下游引物序列(artificial)<400>50cgaaaatggttgacgggac19<210>51<211>21<212>dna<213>h3上游引物序列(artificial)<400>51caagttaagccctttcacccc21<210>52<211>21<212>dna<213>h3下游引物序列(artificial)<400>52tgctcaggatttcaagacgga21当前第1页12