CN104328197B - 红麻表达序列标签微卫星dna标记 - Google Patents

Abstract

本发明涉及一种分子标记技术，具体涉及一种红麻表达序列标签微卫星DNA分子标记。其步骤包括，基于红麻转录组测序序列开发新的微卫星位点，扩增微卫星位点的引物序列以及微卫星位点多态性分析。本发明的特征是，基于红麻转录组测序的转录因子表达序列标签，分离鉴定了24个微卫星标记的DNA序列和扩增该24个微卫星位点的引物序列。本发明的24个新的红麻微卫星DNA标记可应用于红麻品种的遗传多样性分析，遗传连锁图谱构建以及分子标记辅助育种等研究，重复性好，是一种可靠有效的分子标记。

Description

红麻表达序列标签微卫星DNA标记

技术领域

本发明涉及一种分子标记技术，具体涉及一种红麻表达序列标签微卫星DNA分子标记。

背景技术

红麻(HibiscuscannabinusL.)是锦葵科木槿属一年生韧皮纤维作物，具有适应性广、耐盐碱、耐涝、耐贫瘠、抗干旱能力较强、纤维产量高、品质优良及生长快速等特性。其纤维可作为纺织原料、工业原料和生物质能源等，倍受业界新产品新材料开发的重视。

微卫星（又称为简单重复序列，SSR）是由1-6个核苷酸组成的短序列串联重复多次而组成的一段DNA。因为微卫星中重复单元的重复次数不同，所以扩增的微卫星序列的长度呈现多态性。微卫星标记具有多态性较高，重复性和稳定性好等特点，被广泛应用于基因定位、遗传关系分析、品种鉴定等领域，被认为是十分有效的分子标记之一。据序列来源，微卫星标记可以划分为genomic-SSR和EST-SSR，而EST-SSR来源于转录区域，往往跟功能基因连锁（Liu等，2013，PLoSONE8(4):e60346）。在真核生物中，存在着大量的转录因子（transcriptionfactor），这些转录因子在植物的产量、品质、抗逆等性状中发挥着重要作用。因而，开发这些转录因子的EST-SSR标记在性状遗传剖析与分子标记辅助育种中具有明显的优势。

至今在红麻应用到的少数微卫星标记主要来自于黄麻或棉花的SSR标记(Karan等，2013，PlantSystematicsandEvolution，299：619-629)，与此同时，红麻微卫星DNA在红麻品种分类、遗传关系分析中的应用也迅速铺开，有关红麻微卫星标记的专利争夺正逐步展开。但是，国内外红麻的分子标记的研究起步较晚，仅极少数实验室开始了有关工作，目前可查红麻微卫星标记是部分genomicSSR（公开号CN1302111C）。鉴于红麻微卫星数目多，还有大量的未被发现，且EST-SSR具有重要的理论和应用价值。因此，开展红麻的表达序列标签微卫星的分离、鉴定和应用研究，开发具有我国知识产权的红麻微卫星标记显得十分迫切和必要。

发明内容

本发明旨在分离和鉴定红麻表达序列标签微卫星DNA标记，利用9%聚丙烯胺凝胶对这些分子标记进行多态性分析。

本发明实现的技术方案：

所述的24个红麻微卫星DNA标记编号HcEMS047，HcEMS080，HcEMS134，HcEMS199，HcEMS218，HcEMS231，HcEMS270，HcEMS304，HcEMS330，HcEMS363，HcEMS369，HcEMS386，HcEMS398，HcEMS457，HcEMS465，HcEMS489，HcEMS498，HcEMS542，HcEMS569，HcEMS611，HcEMS622，HcEMS626，HcEMS641，HcEMS798所对应的红麻微卫星DNA标记引物序列为SEQIDNO.1-48所示。

所述的24个红麻微卫星DNA标记对应的红麻表达序列标签为SEQIDNO.49-72所示。

基于红麻转录组测序，获得转录因子表达序列标签，利用SSRPrimer软件对其进行SSR位点查找，利用Primer3.0软件设计SSR引物，利用这些SSR引物对红麻8个不同类型材料，即福红992、福红2号、福红航1B、福红B3、福红R-1、KG121、赞引1号与福红952B，进行扩增。通过9%聚丙烯胺凝胶电泳研究这些SSR引物的PCR扩增特点。得到多态性丰富的24个微卫星标记。

本发明的具体步骤是：

(1)红麻表达序列标签SSR引物开发：基于红麻转录组测序获得转录因子的表达序列标签EST(expressedsequencetags)24条，如表1。

表1红麻EST-SSR的表达序列标签编号及注释

利用网上的SSRPrimer工具(http://hornbill.cspp.latrobe.edu.au/ssrdiscovery.html)搜索这些序列所包含的SSR。利用Primer3软件对其进行SSR位点查找，并设计24了对引物，如表2所示。

表2红麻EST-SSR的引物编号及序列

(2)不同红麻品种的基因组DNA提取：采用CTAB法提取红麻8个不同类型材料，即福红992、福红2号、福红航1B、福红B3、福红R-1、KG121、赞引1号与福红952B，基因组DNA。

(3)SSR的PCR反应及电泳：PCR反应体积10μL，反应体系组成为50ng·μL^–1DNA2.0μL，10μmol·μL^–1左右引物各0.5μL，0.5UTaq酶0.1μL，10mmol·L^–1dNTPs0.2μL，10×PCRbuffer1μL，50mmol·L^–1Mg²⁺0.8μL，ddH₂O5.4μL。PCR反应程序为：94℃预变性3min；94℃变性30sec,60℃退火30sec,72℃延伸45sec,共10个循环，每个循环退火温度降低0.5℃；94℃变性30sec，55℃退火30sec，72℃延伸45sec，共35个循环；最后72℃延伸10min，10℃保存10min。Taq酶、dNTP等试剂购自上海生工生物技术有限公司，电泳方法为9%聚丙烯胺凝胶电泳。

(4)SSR的多态性分析：利用上述24对SSR引物对上述红麻8个不同类型材料进行扩增。SSR的多态性和扩增效率如表2和图1所示。发现24对引物得到强且清晰的PCR产物，至少在2个材料间存在差异，表现出多态性。

本发明的积极效果如下：

(1)分离并鉴定了24个红麻转录因子表达序列标签的微卫星标记，经在国际基因数据库中检索，证明为新的微卫星DNA标记。这些微卫星标记的表达序列标签来源于不同类型的转录因子，是具有潜在的理论与应用价值的功能标记。

(2)经将上述微卫星标记用于红麻多态性分析，发现24对引物得到强且清晰的PCR产物，至少在2个材料间存在差异，表现出多态性。多态性分析和扩增效率结果如表3和图1所示。

(3)经将上述微卫星标记用于红麻属的遗传多样性分析，证明这24个微卫星标记适用于红麻遗传多样性研究，聚类分析结果如图2所示。

附图说明

图1SSR标记(HcEMS047)对红麻属8个不同类型材料扩增结果的电泳图，其中1:福红992；2:福红2号；3:福红航1B；4:福红B3；5:福红R-1；6:KG121；7:赞引1号；8:福红952B；M:100bpMarker。

图2红麻品种/系遗传多样性分析。

具体实施方式

下面通过实例对本发明作进一步的说明，其目的仅在于更好理解本发明的内容而非限制本发明的保护范围。

实施例1：基于红麻转录组数据，确定了24条不同类型的转录因子表达序列标签，开发了红麻24个新微卫星标记，如表1，2所示。

实施例2：利用不同来源的红麻种质资源，经PCR扩增和电泳检测，确定出24个多态性丰富的微卫星标记，如表3与图1所示。

表3红麻EST-SSR的多态性和扩增效率

ABCD表示电泳带型清晰等级。

实施例3：利用上述开发的微卫星标记分析红麻种质资源的遗传多样性分析。供试材料来自福建农林大学麻类遗传育种研究室，分别为福红992、福红2号、福红航1B、福红B3、福红R-1、KG121、赞引1号与福红952B。其中，赞引1号是从赞比亚引进并选育的一份优良品系，茎红色，圆叶型，植株高大；而其他材料是课题组选育的高世代品系，茎绿色，裂叶型，植株较高。利用NTsys软件对8份供试红麻材料进行遗传相似系数计算。其中计算方法是非加权聚类分析（UPGMA），读带采取0，1方式，0表示无带，1表示有带。聚类分析结果显示，这24个微卫星标记能很好地用于红麻的遗传多样性分析。聚类分析结果如图2所示，将8份宏麻材料划分为三大类。

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acctaacagccgtgggtgcgtagccggcaatcgcctcaccttgtacgacgttgagatcta720

acggcgaagaatcgatcatgagagcaggctcacggctggaagattccacggtgctactct780

ggctagggctgtggttgttgatgttattaccgttacttttacagtcattccctttgatga840

gattctcacaaggaacaggaaaattagtcttagccttggcgccgcggaattctcgggcag900

cggcgtcgtaggctcgggccgcttcttcagccgtatcaaaagtccctagccaaacacggc960

tcttctttcccggatctctgatctcggcagcgtaacgcccccatggacgcttccttactc1020

ctctaaaatgagcctcattaatattcccggcaccgtttccggttcccttgacggccgccg1080

ttttctccctgggtgccatttgaaaaggaaaaacgaaagcagggcgacgttttaatcctt1140

aatttttggtttcgataatggggattttgtgtggggttgagagaaaaccgatggcaaaac1200

gatccttaatttatagaaagggaggtgtggaatcggcgaggaattagaaagctgagaggg1260

aacacagtcaacgagggatttgactgaagaggggcggctggtagaatagaattaatcggg1320

cggcagtttcgtaaattaagcaatccagattacgtggcgcgtgggtcattcattcatatt1380

atcatcaacggctgttacttgaatccgtgcggtgggtttgggggttgttaacagtggggg1440

aatggagcgcgtgtggtactgatgtgaaatgggaaatgaggaaagtggagttttcaaaaa1500

agctgccggcatattctgctaactcgccgcgtgacattggtttatttttatgttattgat1560

gcgataagtgaggagtggggggctccccggcccgacacacctccttttgaaccgctcgct1620

tggaccaacaggtgttagtggttggaccctattagctgccg1661

<210>53

<211>1323

<212>DNA

<213>HcEMS218

<400>53

ggcgggtttgcgtttcattagattggataaacgttttggaagaagagagagagatgagga60

attgtgaactgtgtggagggttagctcgaacgcactgcgaatcggaccaagccaacctct120

gctgggactgcgacctcaaggttcacggagccaatttcctcgtggctaagcacacccgaa180

ccctcctctgccatctctgtcaaagccctactccctggctcgcctccggtcaccacctca240

gccccgctctctccgtctgcgagtgttgcgtcgccaacaaggacaacgcttgcgatgtta300

cgacggagccggaagagtcatcggaggctgagtacgacgacgatgaagaagaagaagacg360

aagaggcagagtatgatggccttgacgacgacggggaagaagaagatgctgagaatcagg420

tggttccatggtcaggagattcttcttctttctcgatgacgaagccgatggctagctcgg480

attctttgagtagtactgaaggcggcaacggcggcggcggtttgaggttgaagaggatca540

gaagaagccttagtttttgctccgacgtatgttgatttgtttcctttgctctgttttttt600

ttttttttgtccttttttattatattcatctttataaacatgaaattttgccttttcaag660

attgagcttcgaatctggaaatgttcaattaagaatctgggttaaaattgaaaatttctc720

ttttgtttgaaattttcgccatgcattttctaagaaaccaaacagggagttttttttttt780

taacaaggtgaaattgcctctctttttacaaatgcaggatgagattgcctgctcctcgtc840

tcatgtaagatcggtaggatcatctaatggggaagcatcatcatcatcatcaatgagctc900

atcaagactattgaagcgaccaagattagctgaagttaatcagtcggctattattgaaga960

tcagggtgaaagagagtcgagatcaacggctataatcggctctttgaaaagacttcaaaa1020

tcacatgatcatccatgataatgataatgataatgatgcatcagctaccatgattgggca1080

gagatcagagccgttggatttgacctccctctaattctcttattataccctaacctacac1140

gacacatttttttccttttccggaaatgtttcaatttttcttgatatgaaagccattgtt1200

gtttgtttacatttttaatttattttctttcctccttttggtacatttatgactttagcc1260

tcaattgttattttttaaaaatgtgattacattagttttattagaagaggggggaaaatt1320

aaa1323

<210>54

<211>395

<212>DNA

<213>HcEMS231

<400>54

gtttattttggagaaccaccctgcaggtgagctactgtgtcgaacgagattcgatttgcc60

gccgccgccgcccacagctgaacaaggttgctgatttaccaaatactgttgaggttggct120

ttggtatagattctggggaaccgaggcataattcgcctgctgctgctgctgccctgtttg180

attatccagtggcgtttgcgactccattttcactgtgggttctccaaccggtgaactcgg240

cgtcacggtgcaaagattctgctctgtattagcatcaccgctaccagcggcatcattatc300

atcgccgccagacagaaacctggcgacgattcgttccgtctcgggactcgaaggccgatt360

tagaatctcaaggcagaaatccctgtccaagatgt395

<210>55

<211>1206

<212>DNA

<213>HcEMS270

<400>55

ggtaaatggatcgaacttttttccagggtacatagaaaaaagaaacaaaagttttttttg60

ttttcagacagacaatcagacagaaagaaagaaagaacaagtccaataattaatagtaat120

taatacacaacaataatggatacatatatatatatatacgcatatacatacatatatata180

tatacattacatttgcatacaaaagaaggttatgtttacttggcttgagtcattcaacat240

gggcgaatcatccatggtggcaatagcacattggagttgggatgtgctgcagcagcatta300

gcttcatcttctcttccattgctacttgcttcatacgtgcctctgatgttcaaggactgc360

attggttgtggcagaagcatggaagaagaatcttggcagttgttttgttgtgcctgctgg420

acaatttccttctccttctccctctccttcaacttcttcgcaaggacattattctgctct480

tgcaatgctttatcctttttctgaagctcagaaattgattcaaacatgagctgatttctt540

tttgagcgtatgtgcttaagggcagaatcgagttgttgctccaagttttgaagttctcta600

aggctcaaattctcaagatcttcccccatataatgcctttggtttctttgtaaaacctcc660

atcctggctttaagttttgcatgttccagagtccaacttccattttgttcaatttcacca720

gcagcaagttgcctctctgcataagaatacctttcataccgctcgaggatcctctccatg780

caggaatctgaggagaattcaaagagcttccccttggttgagaaaacaatcaaagcaaca840

tcagcatcgcagagaacagagatttcatgggctttcttcagtaaaccagaccttctcttg900

gagaaggtgacttgccggttgatcttgttctcgattctcttcagctgaaccctacccctc960

cccatgaagaacttgaataacaaaactaaaaaagaagccagaaacggagattaaagaagg1020

aaggaaagggattgctatgggaaatatggctatctgtttctctgggtatgtttcatcagt1080

ggttcccttatcatgagttggggcgttggagacaaatgctggggatgaaaagcaggagga1140

gaatttgtgcaagtgttatgtgctgtatgtatatatacaagtttggaaggagagagagaa1200

tatgag1206

<210>56

<211>864

<212>DNA

<213>HcEMS304

<400>56

ggcaatctccgagacttttttttattaattctcattgtcaaatgattcgctcctgaattt60

tgatctttcatttgtcgccacctcatttagactcttccttagtgtttccttgtaaaagcc120

gccgttgcaaatgcctttgactttgtacatttccctctttcaaacacgccattacaccaa180

caaccgattacaaagcagccccccgcctgcccccgcgccctgctctataattttcatatc240

tccctttccctttccttgtacagatttcccattatacggtgccttcaatgttttgttctc300

caagttgctttcaaattatatatgcccgcgatcataaactcaaggttctttctttctttc360

tttctttgttccttcattccttcatcccttcatcgtctctctacctgtatttgtatcaag420

tttacgtttgtctaaatcattttggggtctagctccgggtttactctgctgaattggaat480

tctgggtgtgtctttgtttcttattatctgttgctcaagtctggcttttctaagcgaatc540

ataatctgggtatttctcctattttttatttgttaagacttctgagctgttagagatgtc600

tttgatccataggtcataacctcaaaggtggtttccatttgtttcttcttcttttatttg660

tttgttactttatttatattcctgatcttagagaagagaagaatagttggaggggatcct720

ggatttgtttttgagatggactcaaaaaccccaattgagaatttgtatagtattgaacct780

gccttaagttcctcgtcaagaagaagtaatttttcttctaattccaaatctttgggtggt840

aattcgattagggaagtgaatttt864

<210>57

<211>990

<212>DNA

<213>HcEMS330

<400>57

atttttcccttccaaattttttactgttctccacaatcacaattggatttttcctttcaa60

gtgtttggttttcttccatgaaaattttcgaattgccttaaagttatatgagttcgaaag120

tagtttgagaaatttgagcacttttttttcgaggtaagttttgattcatttcttcattga180

gattagattctgatttggtaaaagatttatggtttttattcggctttgatgatgatgatg240

agtttgatgtcatatttctggaaaatataaaccgacgattgagctgggtagtaaaaatta300

ttggtcaaaatgaacttgaaatttagctctctcttttttttttttaatttaaaacatcgt360

tgtatcatgaacgggttttaattttctaaattaggtaagattttgaatcctgtcatttac420

agctgagaaagttgatatttttgagtaatttcgaagtgtttttgtcgtttgattcaatat480

aaaggaggaaggtagatttcgaatagaaaggagaaaagactcaataggattttagaacct540

tggggatttagtttttctccaaagtgtaagaggcagtttctaggattgtttttggaaaag600

gaaaatgattgttatattgttcttcaagcttgattgcatgggaaagacgaaatttggttt660

tggagctactaaggtgtaggagaagcatgcttttgtagtttttctgtataccattttgtt720

ttatatatatacaaaatttggggtttctaaccgttgagccgagtctttgggtttaatgga780

taacattgaggacgatactagacatcctccaaatccttatggtgttagccatcagcaaga840

ttatggttcttcgagtcaccggaagcttcctgtaagcaatcttccattttctagatcaac900

tgagaatatatccaacgatgacgatgatgatgaagatgacgttgaagatgaggaggattt960

gggaggggaagagaagagtcatgaccaaga990

<210>58

<211>439

<212>DNA

<213>HcEMS363

<400>58

gtaaaactgagatgaagttatcaaactggaaataccttaaacaatcatcaatatataatt60

taaacatttagggaaaaattaaaaaaaaaaaacagacgagaagaaacagtctcactcata120

tatatcaatttctatactttaaagctttgaatcttccgttgattttctctttctccattt180

tttttttaaaattttgtttcatttttagttactaataaaaaaaataaggagaaaaaaggt240

gagaaataatttacggtagatcgagggtgggatgatcttgattactcgtccattgacttt300

gactccaataagtttggtcaacatcgttaggaagatcaaacagccgttgatctccactcc360

cttcccaatccaagggtccgaatccactctccagtccaccatgcaaatttgaaagctcaa420

cctgcaccgtctgatcaag439

<210>59

<211>987

<212>DNA

<213>HcEMS369

<400>59

gcctgacactgctacctctaatcttgcagcacaaggaagggagagtttacttaggaagga60

tatgaacaatctgtcactgaatggactgagaaaactgccgccgggattccggttccagcc120

gaccgacgaggagctggtgtttcagtacttgaaatgcaaggtcttctcctatcctttgcc180

tgcttctatcatccctgacctcaatgtctgcaactttgatccctgggatttgcctggtgg240

gacggaggacgagaggtattttttcagcacgaaggaagccaagtatagaaccgggaaccg300

gataaaccgagcgactgcttctggatattggaaggcaaccggttcagacaagcaaataat360

atcaaggagaaatcaagtagcagggatgagaaaaaccctagttttccacatggggaagcc420

tccccatggatcaagaactgattggatcatgcacgaatatcgtctcgttaccatatcgaa480

caaagatttcgactccctacatgcaaacaacccactaatccagaactatttgaaccatga540

gatggaaaagtgggttctgtgccatgtatttttgaagaaaacaagtaggaacagtagtga600

tgaggaggagaacatgcagagtttttgtaataataataataataatgagaaagtgaagaa660

acctaagacattggttcataagataaaaccaaaattgtatgattttatgagagaaaatga720

tgacatggcagctgagtcatcatcttcttcttcttcttcttggagcagtaatatcacaga780

gatttcttcaagtgcatgcagcagcaggcagtactgaagacaacagccatgcactgcaat840

aatattatttgtataaatcagctgtgttttctctaatgttttgctgtttaatatttatat900

gttattgctttttaatcaagtatttgacttttggaaactatataatataatttgatggtg960

aataattatgagaaaagagagaattag987

<210>60

<211>871

<212>DNA

<213>HcEMS386

<400>60

gagagagagagagagagagagagcagtggttagagagcagagagttgaatcatcgattta60

tttttttttctttttaatttttcgtttccgtcaaaatcggcttctctcgctgtactcatc120

aagatctacctttcatttctaccgaaaatttcgggtttctggagagagaaacaaaaacaa180

atttcaactatttgtttttttttttgttaatttttcttttgaacgttgggatcggaggga240

aaaagaggggttttattaaattttggagggattggagttgtgatattggtggggttttct300

ttttttcgttttttttttgaatatttgttgtaataatttggatttgccgggaactttttt360

ttgttaaagagaattggacttagtttttatgcaatctggtggtggtggtggaggtgggcc420

cggccggaaccctggggtgggtccggttgggcggactgcgtccacatcatctgcagcttc480

gcccacttcctcgtcatcggccttttcggcccctcacttcgggtttgattcggcacgaca540

gcagcagcagcagcagcaacttgaagagaagcaaaaacaatccaaggaaatacttgtgag600

ttttgaaacagggcttaaggttaaaacggaggacttggacaatagagacgatatattccc660

ttcgttttctttccctgttgaatcagaagaagttgaaaacgggttatttgtgtctccagc720

gacatctgaatccaactacttttcagtgtcacctttccacatggggagctttggtttatg780

ccagaatgtgcagacttcggaatccgatctgacggagataatctctgcaccgacttcagt840

tacgaattcgccgatcgtagacctggatatt871

<210>61

<211>814

<212>DNA

<213>HcEMS398

<400>61

gataaggatgaggaagtgggggaaatgggtggctgagattagggaacccaacaagcgatc60

ccggatttggttgggctcttattccacacctgtcgccgcggctcgagcttatgacacggc120

ggttttctacctgcgtggtccctcggctaggttgaatttccccgaccttttagctggaga180

aaaaatcggcgccagctgcggcgacgccgatttgtcggctgcttccataaggaaaaaggc240

taccgaagttggtgctagagttgatgctcttgaggcagctcacctccaacatcatcagca300

ccatcaccgccaccaccaccacctccaccaccaccatcttcatcatcgtaattataatca360

tgattatgacgacgagatgaagccgagtactggtgggtttttacatcgggttgacttgaa420

caagatgcccgacccggaagattcggatgttgtatgggaaaggaaatagtaaatggggaa480

aagggaagttgattgaatcatggccctccaacttttatgattacaaagttgtaatttttg540

gtccgtcgcttttggtggctaccctgagaagaaaggcaggttgttgctaaaatgcaccgg600

gggcaacccctatcgtgcacattgtagtggaggtaattctaggtggcgcagaaaaaaaaa660

atgatgaagccttttgtattaattagtagtttttttttttaatgaaaaatcattttggtt720

tttctcatgttttatttcattggtggtcaagatatattgaaatgtggataattgtataat780

agatgctaatttctaatttctatatttccttcct814

<210>62

<211>876

<212>DNA

<213>HcEMS457

<400>62

tttctttttttttttttttggtttgggttttctgaaaacttggcttcaccgctttccatt60

gcaagggatcaactgattcagcttttgaaaaggaaagtttaaaaaatagaggaaattgat120

atagaaaatggtagagagttgatgtcagtgagggtatttgtagaagaagaaagtggaaga180

tcttttttccagtgcattgaattgatttaaagttaccaataacaagaatggtgggtgttg240

cagctttcgatctaggaaaacaaataattttgttctgaagcagatggttaaagttatttt300

agaaattaggtaaagtgatggatttcaacaattaatatgagcactcctcacaagttatca360

tgaaaaaagggtaaactttttggtggggaaaagacggtagatactgatacaaatgatgat420

ggtgataagaagttaaatggtaactgcaatggactgagtaagagtgtgaaggttagatta480

tatgctttgggcagtgaagttttgttgcaaagttgttcatcatcattgtctggggtggtg540

aagaagaaacagaggtgccaaaggagaaggaagttgggagaagaagaacaagctgctatt600

tctttgatggcattgtcttgtggcacagtttttgcttagaaaccaacaaaataaagtaga660

actaacccataaatctcctctctctttttcttttcattttgtcttgtttgtgaaatggaa720

aaatagccatgtttttttacagctgttctgttttacctttggagagggggagagagagag780

agagtcaaaaggaaagcaaagtttttttttttttgggaatagctgaagatgattgtaaat840

agtactttgggaatgaatggacaatgagaattccag876

<210>63

<211>1438

<212>DNA

<213>HcEMS465

<400>63

gtcgattggaacaagaacaattctattagaagaaaagatatttttaattaatactaaata60

gaaaatagaaatggaaacacaatcagctattggtcgagaacaaacaattggctgtaacac120

aagaggacagcccccattggtcggctgtgtttatctggaatttgagaatgacctaaccaa180

agcatttgtatcgagacaagtggtgacctacctcactcccctcgcctaatctcggctgct240

ctgcctctgcctctggttctcctttggatacataaatgtgtggcgtggggacatttgctt300

ttttctcttttcttttttttcttttcttgaagcttcactgaattctagctaccaaacaga360

ttatggatttacaataactgtacatgcaaaggtgagagatggattcaaaagctatgaatg420

aaatggaaatggaaatggagatcaaatttacttgaggaagatatgatatacttatagcag480

agaaatgtcactggctaggcggcggaggagctgagccagagagtagctcggcgagagcgc540

tcatcgctcgaacttgcatctcaagcgctgctatgtaatctgtagcttcttcgagaatga600

tcggtaacggttcctttcggcaaccgggaactaaccggccgagaactttaactttccttt660

gaacactcggtaaacttctcgctttcaatttcaaaacgctgaatctcggtttcttcgacc720

ggatactaccggagacggcagccacagcggcggcggatcctctatgtctcttccgctttc780

taaacttaagcttaagccggctcgtcagaatggcccggctccacctggttcttcctttag840

ccgccacggccaaagctctatcagcagcttcacggacagctcggccaccgcgtggagcag900

aagaagaagaaggagaaggagaagtgagaattatacggttgctgtgaggaatctgatctg960

tggtggataaagggaaagggaaagggtgaaagtagaagcgaaagagagctagtgggaaga1020

gagcataaagaacataaggcaggaacttagtgatgaaggaacgagagtgagtttggcctt1080

tgagtggactcgttacacccattttttcgaaagaaaaaagattgctctgtgctttttctg1140

ggtaatccaagtctcaaactagaatattctccacctctcgtactgtttttttctttcttt1200

ctttctttctcggtagacaccatagattaatgttttagtaataaaaatgttgttcataat1260

atcaaaattcgaacatctcactgtattcaatagttatcggtttcattaccccacatatag1320

atataatatataggcatttggaatcatattggaaaagaagcagcaaatgccttttccttt1380

atcttttgtattcttttcaatcctaaagatatatattgaagatgaggtcccccggtag1438

<210>64

<211>1678

<212>DNA

<213>HcEMS489

<400>64

ttcccacttcttttttgaaaaatcatttattttttctcttcatttcttgaaaacaaagta60

acgaagtaacgaactctctcaaaatgacttcaaaagttaccaactgttgttgttgtgttc120

cgttaaaaactggtgttgtgattataacggtgttatggcttcttttcggaatttacgaga180

ttgtaactaatatattagaactcgccacaccaactaaagaaaattcatttacatatatca240

aagtggtagatattgcagcaattatttttagtggattaataacaattggagcggcatttg300

gattattcgtatttattttttcaaatacattaaaaatgttattgatatataccaaaatag360

cttatggtatcatagccattaatgtcgcagattatatattatcacttatcgttgctatag420

ttcttaaaccagaagtcatggaacattgtaaaaagtttatttttaataagttatctgatg480

tagaaaaaagtgatgcatgtagtgataattataacgatataataatatctattataatga540

ctgctgttgttacaactcttttcttggtatactttgctatcgtaattacagcctatgttg600

agagtaaacaagctaaagataataataataataataataaaagagagatatgaggatgac660

acttggcatttgtccatttaatgaatgaaaaaacatcagtggccagtcggcggcggagga720

gccgagctagagctaccggctccggagccagataacagatcggcgagagcgctcatggct780

cgaacctgcatctgaagcgccgctatgtaatcagtagcttcttcgagaattaccggcaac840

ggttccttccgacaaccgggaaccaatctgccgagaacctttactttcctttgaacggcc900

ggcacactcttcgctttcaacttcaagacgctaactctcggtttcttcgaccggctagtc960

ctggttacggcagccacggcagcagcggtactggatcctctctgcggcgatctcttctgc1020

tttcggaatttcagcttaagccggctggtcaagatggctcgactccacctcgttctccct1080

ttagcagcgacggccaaggctctatcggcggcttcacggacggctcggccgccacgcgga1140

gccgaaagggaaggaggaggactgagattgatatggctcaaagcttgaagaagcttcgag1200

gaatagatatgctgctgggagtctgatttccaggcggcgtggctcggcggcttttggtgg1260

ctttgttgtttcatcatcgaattcttcttatttttccttttgatacgatcagaatctgtt1320

gtgttggtcagtggattccttatcaacgatgccatgtctcaccggcccaaacgcccctta1380

tccgccgtaccgtacggcgaattatgattcagactgaatggaaaggggaaaaaaaggaat1440

ttagaaattagatacagaattttagataaaatcgaaattctggaaagtaaagagaagaga1500

atttactaccggctgtgaagaacacagatgaaattgagaataaaggagaacgggagaggg1560

agtgaacggaacgggaatgggaacggggtttgttgctttagagagagagagagaaggcaa1620

gaatagaatcagaattgggaggagagcaggcgtcgaatttacaaggagaggcttcggc1678

<210>65

<211>1565

<212>DNA

<213>HcEMS498

<400>65

tcttagtacactcaaacctacaatctttagaagtctacgacagtatttacaagaagctgg60

cgtttataatcaaacgaatagaatatttacaagaagctcatgatgcttgaacagattcca120

tattctgtttttcttttctttggtttccccaattcttttgcaatgttgtatttacaacat180

tctaagttcgtaacaattacttatagttcacaaaatcatgagtcagttggtgtatattct240

cccagcaatatatatatatatatcctagaagagaaatgaaaagaaaaactctattttgat300

cctatgttttgaaatattccatccccttttttttttcccttcaattggactttgctgaaa360

acattcttcctgaaatggctgctgccaatgctgctgtgaaattggggtctttagtcaaag420

aagttgccatctgttccaccaaatgtttcctgacttgtggtgaatccatttctggcttac480

tggaactgctgttcctagcttcaccaccatcactgggcttacattttgtgaggcctagac540

tcatggttggtcctgatggattcaatgaggctgaaccatggcttgaaccggatgctggct600

ccacttgagaaggggggagatggttgtgttcaccttcatatgttgcaaccataactgatt660

gatcatcagcacttctttgcacctgtaaacattaaaacaaacaaactttattcatcttct720

gctccaaattgtgactggaaattgcaaaagtttaaccgtaatgtgatgagacaggcttac780

cttctttttaacagggcagcttggtgcaaaagaacacctgaagtaagctcttggggaagg840

attatccctggtgactttctgcccatatttcctccattggtatccatcctttacaatctg900

gtttcgttttaaccaaattgcatgagaaaataagtaaatttgaaaatcgaaatactcaga960

atccaatgaagtttatatcttaccaggccatcagatgtttcagtcctgacataaactcta1020

gtgactttggcctttataacttcttctctgggtttcttgcatgaatcttcatcggttgag1080

ctgctctccgagttgccaacgatgccaaagttataactattatcattgttgtttccagat1140

tttctcttctttgaagggctaagctccttttctgggtttttgctcattaaatccatcacc1200

tggctttgcaaagcattgtagctctcacacatcgctgataacatttctgtcagcttcttg1260

ttttctgcatttacgcggctcaattcttccgccaaagcaccagtctcttctttcacagaa1320

agtctctttccaaacacaattaagttcccattcggttgttctttcttcggagtttcaagt1380

cgtaggggcctagctgaagaatgattaagatccaaagaagtatccacccaagaagtattc1440

atagcttttttctctcaacaaaactagttcttttcaatcttttaacgaagacccagatgg1500

aactaatccacaaaacacctggaaatcgcctcaaattgaccaaataagaagaagaagaag1560

aaaat1565

<210>66

<211>2308

<212>DNA

<213>HcEMS542

<400>66

taactatgctaccttcactgttaacacgttcctatcctcttctctcctttatttccattg60

tttcttcttcctttctcttttagttctctctttctctctctcttgggatcaaaattctat120

tttattttctctccatttctccacacagccgttttttttcttctttttgccttcaaagtt180

tcaatcttttcacttccaatgcagtctttttgatgtttccttgtgtcgtatcaaccgtta240

ccgagagagaagaaagctcatatttttgaagtttgttttggttttgttttttctcggaga300

aacctcgtgaagtttctggggaaaaaaagcattggtttttattgctttggcgttgtttgt360

ttcgagagaaagaacaagaaaaaaaaaacatgttggatcttaacctcgatatggtctcaa420

gcgagtcatcttgtgatgagaacaccgacaacgacaagaagaagaagaagatgaagacga480

cgacgttaatggaggattcgggttcttcgaaatcgtcggtcatcaacgacgaagatgttc540

cgttatctaatgcaaaagatgagaactcttccaacgacgcttcgccggcgttcgtgttcg600

atatcttgaggaagggaaaagatgattatgccgccaccattgctgaaacggacggaaact660

cgccgccggacttcgttactcgacagctttttccggtcaccggagaaaaagtggggctgg720

agttggagttcgggacaaagggttcaactagtagaccccaatggctgaatttagcttatg780

ctgagtccaccggtgaggccgagctgacgacggctccgataaagcctcagccgaagaaga840

gccgccgtgggccgaggtcccggagctcacagtaccgcggcgtcacattttaccggagaa900

ctggccggtgggaatcacacatatgggattgtgggaagcaagtatatttaggaggatttg960

acactgcccatgctgctgccagagcatatgatcgagctgcaatcaagttccggggagtcg1020

atgccgatatcaacttcagtttgagtgattacgacgaagacatgaaacagatgaagcact1080

tgaacaaggaggaattcgtgcatactcttcgtcgacaaagcaacggattctctaggggga1140

gctccaagtacagaggggtgacgttgcataaatgcggtcgatgggaagctcgtatgggac1200

agttcctcggaaagaagtatatttatcttggattattcgactccgaagtagaagctgcaa1260

gggcttatgatatggctgccatgaaatgtaatggaagtgaagctgtgaccaactttgagc1320

caagttcttatgaaggggagactacaaatagtggaggcaattgtcatcacaatctcgatt1380

taagcctcgggatttcagcgccttcggttgctcgaaatggaagccatgatgctgcagttt1440

cccattttcgtaaccctgctggcaccagagagaggctgcagtttgagagtcctgtttctg1500

cgtctgtgggcgtacaacctcaacatggtccaacagttgtagccaagcactctccgatgc1560

tatccggtgtttacccgcctgctcttttaccgtacattgagggaagggcaatgccgaaga1620

gagccgaagctgttccgtcgaccggattctcgaactggggctggcaaattcaaggggatg1680

gtaaagttacccaaaccccagtgtttcctatagcagcatcatcaggattctcttcgtcca1740

atgccattgcttcatcgactactctcccgttcaacaattcgggccgaaatctttgccttg1800

caacaccgactacacccacaaataccacgtctcattactataattacaggagctgaaatc1860

ggcacacgactccgcgaaaacgttccataatgcctcaccgtacaagttaggtgagtagaa1920

caagtgagagctgtaacaattgtggtaaatccatggagtttgcattcgttacaaatgtgt1980

aatacaaagaatacattgcatttgttaattttgcaaaacttaaaagttctcgcctttgta2040

acgctttttgcggtcgcgatgtcacggttaatgttttgtagttatgacttccgatatcgc2100

tgcggaaatccgaataacatcgcggtgttagccgagtcggactaatccgagatgcaaaga2160

aagacatctccggaacagggatttggaaatggaagattttttagtagtgtaaagaagcaa2220

agcaaattatagttgagaagaaaaatgatgatgttgggatccgtcggcactcccatctga2280

aaggacttgataactcgtccacttatcc2308

<210>67

<211>565

<212>DNA

<213>HcEMS569

<400>67

cagcagcaacaacagcagcaacagcaacagcagcagcagcagcagcagtagcagttcgag60

tagaaatttaagcgataatttgaagcagaggctattatctctgggaagacaagttttgat120

gcaaagatcgacggtcgagaatcaaagaaccaagctaggagaagaagaacaagccgctgt180

gctcttaatggcactctcatacggctctgtttatgcataaattcctttctttcttttccc240

tctaatatatatgaattttagagcaagaatgataggttctttaatggggttttctaacct300

ttcatgtgtaatcaatctctagtgttttagtctctacttctttgttttaggtttaactga360

caatggaagagaagaatcccaggataaaaaaaagtggggatttttcactccattttccct420

aatctcgtattgctagtaatttgaagttttagggatttttatttctctattttgtgttca480

gtttatttatattaattattaattgttttatggggtatgtggttttcatattatatgtac540

tcggtcaaatcaaaatgatgtgaag565

<210>68

<211>3362

<212>DNA

<213>HcEMS611

<400>68

ataaaactaaattctggtatgttgtatttgtgatttttgatctgatttgttgtcaaatgt60

acaacaaattgtcaccaccttaaccagaaaacgaaacttctcactgaaaaccaaactaaa120

caagaatacgatagcagcaacatcaatagtccaaaacgaaaagcccatactagagttttg180

cagtcagatgttttcttctacagctactgcctactgggaattgcaactatactagacaaa240

caagagcggagaagactagcaaatacacgaaaatgaacaagtactaagtcataaaagctg300

cagatattaccaattattgacttaattaattcttctggatagagttgatcgattgactga360

caaataggacaatgaacgcgtactaagtcactgccccgtctcggaggctgctactcagca420

tctccaagctgaagctgagataagatccctctgctgccagcaaagaacaagtgatgcctg480

gcgcttttcaacatgaaatcccctcacttgagccctctttacaacacaatctgcctgagt540

ttcagtgaagttactgaatgtcacaggggttaaaccagccaaggaaaaaagcatcttcca600

aagaggtatcttctcaggtgcttgtagtcgccccaacacagtgctttcaattctcggcac660

aagaaggaacctctcgatcttgttcacagcatcggatgtcacgttactggcatcaagcga720

ttctaacaagtttatatatgactggaaggcatggattacatgttgtgggaatggcagatc780

atttctatcacacccttggtctaaagacacaacaatttttggtgaaagctgcttcacaaa840

gtgcataagattaggtaaagcagagggccgattcgaagaagaccaaacagggagattcac900

agcaactgcctcgttttcatttggtcggaacattggcaaagaataaggggtctgatctaa960

agaatcaaaattaagcacctcaagctcaaaacttataccaatttcattagcaaactgcac1020

aagattttcccgcataagctgtagctcaattggatggtgtgttgaaggtgaggcaaaggc1080

agtaattctcaacgacagcgctcctctactcctcatgggaagctcctgcataaaggaagc1140

ccactgagcaccaaatccaatatcaaagtctacaatgtgaatccgatcagtatcatcaag1200

agcttcgaggagagcctggttgcaagtaaaattgacgaattggataagcggagagacttc1260

ggagaagaccttgtaagcccccattttgaagataacatcgaaaggggtagggctccttgg1320

aggtgggggagagactgggttatgcatgagaagtaattgcaaagcctccttgaagtagaa1380

agcagccctttgtgggggcatacctactggggagacctggtgattgagccgcgccaatat1440

caattgcgcctgtgagaatttcccagtcccaaccaactcagcagccttgaaaagctggtc1500

gaataatgtggcctgttgatgctggtgtaactggaactgatgctggtgttgtgctgtctc1560

ttctcccgaacccagcactttctgtttatcgaccatcagcgactgctgctgtgtaatata1620

ctggactccctgagaaaaccccatgtgctgttcttgctgttttctcataaacaactcatg1680

tccggcatcagaaaatgggagttttgggatctgataagaaagttctaagttaccaggatt1740

aaggcgtttcggtaaaggctgaagtagctgttgttcttgaggcaatggcaagaaaaagtt1800

tggattttggggaagctgttgctgaccaatcaaaatctgaggtttcccttcttggttttc1860

aagatattgaacttgctgctgctgttggtaaagcataggcaaagaatcaggaagaccaaa1920

attgctagccggagaagtaaaggttggattttgaataattgtattggaccctacagtagc1980

accgagaactttgcgattgttaaccagctcaactgaagagctttgaatgaccgaacccat2040

ctttccattctcactactgttggcattcggcagaaaccaagaaccgggaaaaccacccaa2100

attaagagcagcacttgaaatcgcgttaccaggaccagaacaagcaaggttaccaattgg2160

gtcagaccctggaccctgatcaacaacccccgaccccgcattgccttcgaaatcaagacc2220

attacttccaccatgaagcagctgcttgagaccgaaagaacgcagaagcgagtggtcttg2280

attcagaaacatggctgaatcagacatcatattcccccaatcttccaatccaatactgca2340

tctttcaacgcctccgggccctggaacaaggtctagttcaatcgggatgggttgaagctc2400

cgtcgcccattcttcttcgttgttgctcggaactgtactctcgagtggtgcaatggcacc2460

cgccgtggtgttggtgtttccggttgaattaacttcgccgctagcaccggcgttgaaaga2520

agaagagagagtggacgcagatgtgggtgggctttggcttcttctcatatgaagaacaga2580

tgtcggttcttgttcgttgttgtagaagaagttattagcttcttgctgctttacccactt2640

gtcttgttgacaaatcgaagcaaactccgtaagttccaccgccgctttagcctggaactg2700

aaagtgcatttgtattatcgtatacaaacgaaaagacaataaaaaaacgaagctgttttt2760

tttttcttttggagaatcagaaattaaggagaaagacaacttttttttcctacaacgaaa2820

aaaaaaagagagaaactttatacactgttcttcaagatttacgaaacccacactatcttt2880

ctttgtatgccatgaaatcaatctcccaacttcaatagaaaaagacccattttgccaaaa2940

tagctccttttgttgttctttctctttctcctactctccatctttctttttatctcctca3000

actcaaccttaccaaaactccatttttgtagctccatctctctaaccagctctgttcttc3060

ttctttttttttttacttttatctctctccctggttggaatttgtggctttatcgccggt3120

attactgtaatttcccgttgttatttgctgctaactagtacctcttatgtcgtagcagta3180

ctctgttctgccatggatgcgtgcatcataaaaaaggcaataaagtactctccctcttct3240

ccatctctctctctctagagtctagtccctattctctctctctcctctttgaatatattt3300

ttcgtttctatttttcgctaattttttatgcaccacctctctctatctctctccacatct3360

ca3362

<210>69

<211>2294

<212>DNA

<213>HcEMS622

<400>69

aagaaaaggggctcaatttatttgacaaaaggtggtaaagaactaaagataatggatgca60

tgagccaccaccaccttcttggaaacactcttttcttctcacttctctccccatctccca120

tttaccaacaatatcctcaaaatctgccctttctttctttctttctttcttttttgaacc180

caaccttccctacttcacgtggctacgaaagtagtctctttcttactactgaaatttcaa240

tttagcaaagttacaattttatatacaatatttttcatatgatgcaaagcgtcaggcgac300

aacattgcgtgacatcctgttagaaaaaaaacagaatgggatataggagaaaaaaacaaa360

tggcaagatgaattcatcttctttccagaaacgaatcatactaaaaacctcttattagtt420

ctcaaaacttcacaacagattggcaaattcctgagatcagatataaattgcggaatattt480

gtccataaaaatgaaaaataatgttaacacaattgataaagcggctgtcagggggttggg540

gtgaagcttcactaacatgccttatttacaaatgaaacaaagtagaactagagcatgtct600

tctcagaaaagaccaccagtcatggaagagaagtcatcaaagctccaaaaatccattggg660

tttccaccatcctgagttgcatctccattgagaaaggcgtcaatggaggcatcccagttg720

ccccaggtcatctgatcgtccaaacccaatagctcatccgacaaactcttcccagagttg780

tcttctgtaggcataacgttgtcagaacttggtttcaacttcttctgagggtttgcatcc840

tccagaaactcatcgctttctatagaagcttcaggaacagacgatatttcaggagtcttt900

gcaccaagttctccccagccgaagtcagaacagtccaaggagttgcttccatgatcagaa960

ttgaaatacaagggagagttgtcagattgagcaaagggtttagatccaacatctacaaca1020

gcaggaacagggtccacgtgtgcaaatggatctatcggtgccttctcctcaacgaaatcc1080

atgatatcatagtagttctgctctggtttgctcaagcaattgaaattctggttgatattt1140

ggctcaacagggctcaaattcgacatgggaagtggtttctcatcatttttcttaatcaca1200

cgctttggagaggcatgcgaaatctcatcagggaagttcacctttgctttcataccccga1260

attctccgtgcctcagaatcatatgctctagcagcttcttcggccgtattgaaagttcct1320

agccagaccctaacccctttccttgggtctcggatctcagcagcccatttaccccaagga1380

cgttgtcggattcctcggtactgggtcttcttctttctttttgcacacctttcagcttga1440

cctttggattgcacagattttacagggttcggacctgcataattgaaaaatagaaaaaca1500

tattaatattgtactcattcacaaaccttacgaataaatgttatagagttaataaaaaac1560

tataaatcattaaacaactttcaccgggcaacagaatttaggaccttcaagtgcaagaaa1620

atcatgagaagaatatggtagtaaatattatcctcaaaactcgagactcttggttaaggt1680

atggcttctcaaactcaattccccaattaattaaaggtaaaaagttgggtttctcctttc1740

agactcaataaacatgatcttcctccatactaagatttcaaaaaccagtttgacacaagt1800

gaaaaggaaatctaggaaccgtaggaataagtcagacttaaacaataagaaagaatcaaa1860

ctgtatatcaatctcaagagcaaaatcagttgacttgatcatgtataaaagccactcttc1920

ttacagatcttcatttattcaaattgctaagattttatgatctaaaaagtactactccta1980

gtaattttttcttcttatccttctgtttacaaacaaatctcaaacagcacaggaaaataa2040

ttttctaaatatataaagcatagaaatcagcaggaaaaagcagatatgaagggaaatctt2100

ttatacacaagcaaagacaaaatctaccaacgtctggggaaaaaaaagaaaagaaaacag2160

acaaattccataagtttaccttcaattttttttcctacaataaggagtaaaacaaacaca2220

cacacaaaataaacaaaccaaatctgaaagaacaatgaaaaaaaactaagaattcaataa2280

tattgttatcatat2294

<210>70

<211>1168

<212>DNA

<213>HcEMS626

<400>70

ctcctctaggtgtgcctccattttcatgaaccccactttttatattcatcatattgtatt60

tcaccagacatgtttgcttccagttctgtctctctgtgagtttgttatctgccggttgtt120

gttggtctgagatttgggtgggaaagctttgtcaggaaggaaatgggcaggaagtgctca180

cattgtgggaacataggccacaattctagaacctgtgctactttcagggatactggtggt240

agtggtggtggtggtggtggtggacttcggcttttcggtgttcagctgcaactcgattta300

ccttcgtcgtcgtcgtcgtctgttgctgatgtggttaatcatatgaagaagagtttcagc360

atggattgctttccttcttcttcggtctcttcttctccttctccttggtcctgctctcga420

gtttcggtcgatgagaattccgataaaatctctgtcggttatctatcggatggtctcatc480

ggccgatctcccgagagaagaaaaggagttccatggacagaggaggaacacagaatcttt540

ctaataggacttgagaagctagggaaaggagactggagaggcatctccagaaactttgtt600

actacaagaactccgacccaagttgcaagccatgctcaaaagtattttctccgacaggcg660

acgctcaacaagaagaaccgacgttccagcctgttcgacatggttagaaacagtagcatg720

gccggacctcctcgattcgactcgaatcgtcatcgacagcacgagttcaatgcagcagaa780

tccatgtctccggcgcccgatctcgagctcacgctggccgccccgaggccggccttagaa840

gaaaacaagtcatccccaacaaccctcctgatcagacctattagtgttacctgaacacaa900

tactgtagtaattagtagtaattaattaattaatcaatctttccttcgttaatcataatt960

agtagaatcttcctttcattctttgtccataaactcagttaattacaatatctggagttt1020

gtgacctattttcataaatgccctttagttccctctcttttagcattatattagtgtaat1080

gtgcaagtgtaatcatttggtggattccgattatatctggagttgagtcggaatttagcc1140

tttaaatgaaatagtttttttctactca1168

<210>71

<211>1801

<212>DNA

<213>HcEMS641

<400>71

acaaaataatatacataaaacaatattaaaatgccaacaaaaatcaagaaaatatctaaa60

aaaataaaaagaaatcaacaattaattttttttgacaaacaatcagacatatacaccatt120

gtctctgacaacactaactcttttcaatttcatccctcccaatacaaccacacgccttaa180

aatcccaccatgttttgcccccaaactaattacaaaaaaaaaaaaaagttgaattatttt240

catcataataaattggaaaaaaaagaccaattcccaaacaaaacccacacaataatatta300

attaaccaattttaaaaaaatgaaaaaagaattaaattgtcaccctcctatcttagcatc360

ccacgtgtcccttttttagcccgtcaattttttttttgtctaaaagggacacccctacac420

tagtagaaaaacagcaattgggaaacacataacatgaaaaaaaaaaacagccccccaacc480

ccattgttagatagctttattgcattattttttaatttatttttgcaccagatcttctcc540

ctgccatttctttgatcaactcacgagttgttcttcattctccggcggcggcggttgttg600

ctgcggtggtgcatggtggctgttgttattgtcgttcattttcttgatgctttgctgttg660

ctgatagacttgtcttagcctctcaatttccttcttcaatgcttcttgatgagcatcttt720

gaaaatcttgtcttgagccaaagctgcaattctttgcttcaaagcactattgtcaacatt780

gagaatcagtctttgatgatccaagaatgcaacccttggtgataatgctgatacttcggt840

ctgtaatgttgtgacgctcctttcaagctctgaaatatattgcagctttctcactcttga900

tctttgtgctgattgccggtttgccagaattctgttcaatgagagggttttaaaatggtg960

gaaaaaataatggggtttgattttttgttttcttttacctcttaaccctcttcggatcga1020

cgatggtgtcgccattggagttggaaggaggctgagcagcttgtggttcagatttgcatg1080

agctctctacttctcccggttcattcttggggtgttgcagatccagggacggcaccggtt1140

tctcgtcgttgttgctgttttgatccgacgtgggcgtcgaagggttcgacgaggatacgg1200

tcggggcggcagcagccatggcgacggcgacgtcgtcggagaacatagacatgagctgct1260

cgtcgtcgaggcggttgaacacattggcatccgtcatcatggcattgttgttactattgt1320

tgttggtggagtccctacattcttccatcatcaacggttgctcgaggaaggcgatggagt1380

cgctgatggaccgacggtgagccccgcgacgcgccgacgagaagtcgaggaactcatcca1440

cccatgaaggctggtggtttgagtcattcatggtggtgggaatgaagttgggcatggcgg1500

gcaatctttggtgcgggaaagatgaccaattttgggtcatgttgggtatctttggaggta1560

attgtgccatttctaaagctaatgtcacaaaccccaaactcaaaagacaaagcaaaagag1620

cacgaaaaccaaagctaatcttaatcaacaaaaaaaaccaaccaactaactgtacaacca1680

acacacaaacatgtcacaaaatcccaacatatatcaaaacaccaccgaaaccgaaaccga1740

aaccgaacacaagtatcatagaatgataagagatagttgttgtgcaagcaaatgatataa1800

a1801

<210>72

<211>1142

<212>DNA

<213>HcEMS798

<400>72

tgcagttgtaatagcatccaacatgattataacgagaatttgttaaagccgacaacaaca60

aaaaagttgtacatgttaaaattagagttgagtatactacataacatgatgactatagat120

ttcatgaaccaattaaattaaacttgaggttcggattattattatcttgtaagccatcgg180

catccattcttcggctcatcattttccttctatgaatatccgaccaatcagtataactag240

ccctagttttgtaacatgtctcttaatttggacaagcagattgaccggggtaaaatatta300

taggtggcatcctcaagtttgtctcgcagctgaaacactgatggtgaggtccaggttcaa360

atcagggagctgattactctcgtccaagcagctttcggcatcggaaacccgatcgttgtc420

gcggtgtcggaagttagccggcggaggaatattatcatcaccgataacttgaactttcgg480

accggataatatttcagctgcgccggtatcggaattacgtgggatctgagggcgaggaag540

cagattgagagtcgttaaccgatggttattcggatcgacacccatgtttacaagcttcct600

tcttagatgagagttccaataattcttcacctcgttgtcggttcttccgggcaatcttcc660

ggcaatcaacgaccaccggttgcctaagagtgcatgaagcttgataataagatcctcttc720

atcttctgtaaagttgcctcttttaaggtctggccttagataatttatccatctcagcct780

gcaacttttcccacatcgaaggagccctgcagcttgaggaagagtacgccagcaaccttc840

accatggattcgaatgtaatttacgagcttctcatcttcttgcttagaccaagccccttt900

attcgtgtcttgcttagcacagcaaggcttcctcattgtgatctccaccgaaaacgcggc960

cgcaatcgtaatttaaatccccggatacgattacagtcggataaaaagggaatagaaagt1020

atatagcaaggctttttcttgttgttgttgttgttgttgctacataaaatattgaataat1080

gggaatataatagatgaagggaaaaatgaggaggagggggttgagttgactggtggtttt1140

gg1142

Claims (1)

1.红麻表达序列标签微卫星DNA标记的引物，其特征在于：所述的24个红麻微卫星DNA标记编号分别为HcEMS047，HcEMS080，HcEMS134，HcEMS199，HcEMS218，HcEMS231，HcEMS270，HcEMS304，HcEMS330，HcEMS363，HcEMS369，HcEMS386，HcEMS398，HcEMS457，HcEMS465，HcEMS489，HcEMS498，HcEMS542，HcEMS569，HcEMS611，HcEMS622，HcEMS626，HcEMS641，HcEMS798，其对应的引物序列为SEQIDNO.1-48所示；

所述24个红麻微卫星DNA标记表达序列标签为SEQIDNO.49-72所示。