CN104328197B - 红麻表达序列标签微卫星dna标记 - Google Patents
红麻表达序列标签微卫星dna标记 Download PDFInfo
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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
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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ccaagcttgtgtttgtgcttgtgcacccctccacctatgttttgtttttgtttttgtgtg180
tgtgtgtgttttttttttgggttcatgttcaaattacataatttacatgtttttttagtg240
tatttcatcatacgaggggtgtacggtaaagaagaaagaaagaaaaacctaaaactaaga300
agaaaataaaggaaaagaagatccgaaattcatgatcttcatgtgtacacgtgcgattca360
agcattgcattccgcccatccttcttccaggttcggtcttattctagtcttcctcaacct420
catcgcacgtgccagcctctgctccacgcccaccaccactctacccggcacgcccgacag480
ctcctccaccctcctcaaattccgccccgacgctaccgacgtcgtcctcagccccaacca540
aaacgcagc549
<210>51
<211>862
<212>DNA
<213>HcEMS134
<400>51
gagaccacaacaacgcccacaaatagaagaacctgtacaaaaactctgcccacggatccc60
ctttttattattattatttattaccattttaattattgtttattaggggaatttccaaag120
gttgtcagcatttccatcgttatctgcgggaggcggataggtagcggcggcgtccatgcg180
aggagggctaagcagcatcccctccgccatgttgcaaagaacattcggcatatcaaatat240
caaatcctcgtccacgaactcatctccccttgaatgcctcccacccgcctctactgttgc300
attgctgttgcttctcgttctcaaagcatcattcgccgcacccaaagccgacgccgccga360
ggacgcagccgcctggatgtcacgcggggaagtcgacgccggtaccggcaacgaagccac420
agagttggggaagttaacctcggcgtctcggccctttaaagcaagggccgcaacatcgta480
agcaatagccgccatttcagcggtagggaatgaaccaagccagatcctgttaggcttcct540
gggctccctgatttcagacacccatttgccgttactcctgcgcctcactcccttataaac600
cgggtgccgtccagtagcggcagaagcgtcgccagtgctgctgctgctgctgctgcaacg660
atggatccttgagtactccatttcttcgtgcacggtgaagcacgatgatggggacatcat720
atcgcaaacggagaaacaagacgatgcccggcgggcgcggctgatcagatccctttccat780
ttcggacatattggtttcttctttgcctcttacttgtggctagatttgattttatgtttg840
aagttgatgaatgatgtgtttc862
<210>52
<211>1661
<212>DNA
<213>HcEMS199
<400>52
gaaagaaaaaaaccctaattaaaatatatcagcaaaacacgtaaagtcgtaaactttaac60
ttcaagaggattatggataagaattgaacaaaatggaaacaaatgaaactttgtagagct120
taattcatgggaaatacagaaactactgatgaaacaacagagaatctgagatttctctgc180
tgcaaaaaagtgtagttcttcagctaaactatacacataatctgggttgtcaccgtagat240
cacatcaaaatatgccgtaattttttctcttttttttggtgtctcgttttttttttcttc300
taatctaccttcgattcagcggacggaaaataagaaatttcaagcttttttccagatggg360
cgatcggcgaatattaaaaaaaaaataatcaggcgatttcaggaagagccggctggttga420
gatcgatgttcagaagcgggcgagacttgacctcgcggtggttcaaatcgacgatggacg480
atgaatcggagtcgctttggacaccgccgttaaacgccgacgtgtggaaatcacgatgat540
gatcgaatctcaaccgttgaaactgctgaccgttcactacacccggccgaacgaagggat600
caaagtaaaagaccttgccaccggcgatatacggcacaagcgaagcttgctggaaaggaa660
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。
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