中华绒螯蟹核糖体DNA全序列分析
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中央级公益性科研院所摹本科研业务费专项资金项目(2007JBFB10);农业行业专项优质蟹种规模化繁育与养殖示范(nyhyzx07-045);农业部水生动物遗传育种和养殖生物学重点开放实验室开放课题(BM2007-04)


Characteristics analysis of complete ribosomal DNA sequence in Chinese mitten crab (Eriocheir sinensis)
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    摘要:

    核糖体DNA通常被选作系统发育研究的分子标记,核糖体DNA中IGS部分序列的变异在一定程度上影响着生物的生长。报道了中华绒螯蟹核糖体DNA全序列的分离和序列特征。中华绒螯蟹核糖体DNA全长11 660 bp,包括18S(1 873 bp),ITS1(317 bp),5.8S(163 bp),ITS2(614 bp),28S(4 461 bp)和IGS(4 240 bp);不同部分AT含量在40.1%~48.6%之间,低于果蝇(55.8%~80.0%),高于鲤(22.0%~43.7%)。平均100个核苷酸含简单重复序列(SSR)数由高到低依次为ITS2(0.98%),IGS(0.49%),28S(0.38%),ITS1(0.32%),18S(0.21%),5.8S(0%)。个体内差异分析结果表明,中华绒螯蟹个体内存在两类ITS2序列,其中一类在405 nt处有一12 bp(CGCAGGACCACC)插入序列。中华绒螯蟹rDNA的IGS部分长4 240 bp, AT含量为42.6%。IGS部分包含一个有约7个连续136~139 bp单元组成的重复序列区,重复序列单元中存在XhoⅠ内切酶位点,为河蟹特有的重复序列;重复序列区后有一个由182个碱基对形成的柄和21 bp的环组成的发夹结构。中华绒螯蟹核糖体DNA序列的揭示将为继续研究不同地理种群间的差异标记和挖掘与生长率相关的核糖体DNA分子特性提供理论基础。

    Abstract:

    Ribosomal DNA is usually selected as molecular marker for phylogenetic study and the sequence variances especially in IGS region can affect the growth rate of organisms to certain extent. A complete rDNA of Chinese mitten crab (Eriocheir sinensis) was isolated in this study. The rDNA is 11 660 bp in size and consists of 18S(1 873 bp), ITS1(317 bp), 5.8S(163 bp), ITS2(614 bp), 28S(4 461 bp) and IGS(4 240bp). The (AT) contents of different regions range from 40.1% to 48.6%, which are lower than those in Drosophila melanogaster (55.8%-80.0%) and higher than Cyprinus carpio (22.0%-43.7%).The SSR loci per 100 nucleotides in the sequence of descending are ITS-2(0.98%), IGS(0.49%), 28S(0.38%), ITS-1(0.32%), 18S(0.21%), 5.8S(0%).The results of ITS2 intra-variances analysis showed that two types of ITS-2 existed in Chinese mitten crab, with a 12 bp(CGCAGGACCACC) insertion at 405 nt. The IGS region is 4 240 bp in size and there are 136-139 bp speciesspecific repeats that contain an Xho I restriction site. And a stemloop structure (182 pairs stem and 21 nucleotides loop) is located following the specific repeats. The sequence of rDNA in the Chinese mitten crab will provide the foundation of studying the variances among different geographical populations and exploring the molecular marker associated with the fast growth rate of the Chinese mitten crab.

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俞菊华,李红霞,戈贤平,李建林,唐永凯.中华绒螯蟹核糖体DNA全序列分析[J].水产学报,2010,34(5):696~703

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历史
  • 收稿日期:2009-10-20
  • 最后修改日期:2010-01-19
  • 录用日期:2010-03-08
  • 在线发布日期: 2010-05-10
  • 出版日期: 2010-05-15