凡纳滨对虾EST微卫星标记初步筛选
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国家“八六三”高技术研究发展计划(2010AA10A401);广东省海洋渔业科技推广专项(A200899A02 & A200901B03);广东省/中科院省院合作项目(2009B091300088);中国科学院知识创新工程重要方向性项目(KSC-XZ-YW-N-4701)


A preliminary study on microsatellite markers screening from EST sequences of Litopenaeus vannamei
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    摘要:

    对161 075条凡纳滨对虾dbEST(database of “Expressed Sequence Tags”)的数据进行SSR序列筛选,搜索参数为二~六核苷酸重复6次以上(包括6次),共获得含SSR的EST序列12 600条,占整个凡纳滨对虾dbEST数据库的7.8%,其中二核苷酸重复10 104条,占总ESTSSR序列的80.2%; 三核苷酸重复2 036条(16.1%),四核苷酸重复336条(2.7%),五核苷酸重复35条(0.3%),六核苷酸重复89条(0.7%)。大部分发现的微卫星序列为完美型(perfect)的重复序列。在二核苷酸所获得的SSR重复单元中,AG/CT是优势重复单元,共分布4 820条,占二核苷酸各重复单元的47.8%;其次是AC/GT重复,共分布3 584条,占二核苷酸各重复单元的35.4%;最后是AT/TA 和CG/GC,分别占二核苷酸各重复单元的16.6%和0.2%。通过对凡纳滨对虾EST序列中SSR位点信息的发掘分析,共设计77对微卫星引物,在10个个体中成功扩增的引物有54对,成功扩增率为70.1%。其中多态性丰富的引物28对,多态率为51.9%。等位基因数为2~5,PCR产物大小为140~600 bp。本研究筛选的ESTSSR标记将为凡纳滨对虾的分子遗传学研究、种质鉴定等提供可靠的工具。

    Abstract:

    EST-SSR marker development by screening and mining the SSR repeats in the EST database of Litopenaeus vannamei,161 075 EST sequences were downloaded from EST sequences in the database of GenBank.EST sequences were analyzed and the SSR was screened with SSIRT(http:∥www.gramene.org/db/markers.ssrtoo).As a result,12 600(7.8%)SSRs were identified from the EST resources,among which there were 10 104(80.2%)dinucleotide,2 036(16.1%)trinucleotide,336(2.7%)tetranucleotide,35(0.3%)pentanucleotide and 89(0.7%)hexanucleode SSRs.Among the dinucleotide sequences,AG/CT repeat motif accounted for 4 820(47.8%),3 584(35.4%)AC/GT repeat motif,AT/TA,CG/GC repeat motif was 1 673(6.6%),27(0.2%)respectively.77 pairs of primers were designed with Primer Premier 5.0.Fifty four primer pairs obtained expected product,with the success rate of 70.1%.And 28 of them showed polymorphism among 10 individuals,polymorphism rate was 51.9%.Number of alleles ranged from 2 to 5,and PCR product size was 140-600 bp.This implies that the polymorphism ESTSSR developed from NCBI could work in genetic mapping,gene clone and gene functional research.

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王艳红,胡超群,张吕平,夏建军,任春华.凡纳滨对虾EST微卫星标记初步筛选[J].水产学报,2011,35(7):969~976

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  • 收稿日期:2011-01-13
  • 最后修改日期:2011-03-28
  • 录用日期:2011-04-20
  • 在线发布日期: 2011-07-13
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