文章摘要
陈合格.中华鳖与砂鳖线粒体DNA 12S rRNA基因序列的比较分析和分子鉴定标记[J].水产学报,2005,29(3):
中华鳖与砂鳖线粒体DNA 12S rRNA基因序列的比较分析和分子鉴定标记
Comparative analysis of mitochondrial DNA 12S rRNA region between Pelodiscus sinensis and Pelodiscus axenaria and their molecular marker for identification
投稿时间:2014-03-13  修订日期:2014-03-13
DOI:
中文关键词: 中华鳖 砂鳖 12S rRNA基因 PCR-RFLP分析 分子鉴定标记
英文关键词: Pelodiscus sinensis  Pelodiscus axenaria  12S rRNA gene  PCR2RFLP analysis  molecular marker for identification
基金项目:农业部中华鳖标准项目
作者单位地址
陈合格 湖南师范大学生命科学学院 湖南师范大学生命科学学院
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中文摘要:
      对中华鳖和砂鳖线粒体DNA 12S rRNA基因进行了引物设计、PCR扩增、序列测定和PCR-RFLP分析.研究结果表明:中华鳖、砂鳖线粒体DNA 12S rRNA基因片段的碱基序列长度相同,均为562bp,其A、T、C、G含量相似,分别为209个(37.2%)、121个(21.5%)、145个(25.8%)、87个(15.5%)和207个(36.8%)、120个(21.4%)、145个(25.8%)、90个(16%).两序列间共有13处碱基不同,序列差异率为2.31%,中华鳖与砂鳖各自个体间的平均核苷酸序列差异率分别为0.53%和0.36%,种间差异显著;用内切酶MspI酶切两种鳖的12S rRNA基因片段,在砂鳖中可得到大小为519bp和43bp两个片段,而中华鳖无此酶切位点,这可作为准确鉴别中华鳖和砂鳖的分子鉴定标记.从两种鳖线粒体DNA 12S rRNA基因片段碱基的显著差异和MspI酶切位点的变化,可以进一步证明砂鳖是不同于中华鳖的鳖属一新种.
英文摘要:
      The primers for Pelodiscus sinensis and Pelodiscus axenaria were designed by using the sequences of mitochondrial DNA 12S rRNA region of Dogania subplana , Chrysemys picta and Chelonia mydas , 562 base pairs of mitochondrial DNA were amplified and sequenced. Theresultsshowthatthelengthoftheirsequenceisthesame,theA,T,G,Ccontentsaresimilar,thenumberofA,T,G, Cis 209 (37.2%) ,121 (21.5%) ,145 (25.8%) ,87 (15.5%)in Pelodiscus sinensis and 207 (36.8%),120 (21.4%),145 (25.8%), 90 (16 %)in Pelodiscus axenaria. Based on their sequence data , we found there are 13 different nucleotide sites (percentage divergence is 2. 31 %) between Pelodiscus sinensis and Pelodiscus axenaria by using the software of ClustalW , whereas the average number of different nucleotide sites is 3 (percentage divergence is 0. 53 %) in Pelodiscus sinensis and 2 (percentage divergence is 0. 36 %) in Pelodiscus axenaria. The nucleotide percentage divergence is obvious between species , so it suggests Pelodiscus axenaria is a new species in Trionyx. Restriction endonuclease analysis based on sequence data of this DNA fragment revealed the presence of polymorphic sites for MspI endonuclease , there is one site for MspI endonuclease in Pelodiscus axenaria , but none in Pelodiscus sinensis. The restriction profiles obtained by agarose gel electrophoresis when amplicons were cut with MspI enzyme allowed the unequivocal identification of Pelodiscus sinensis and Pelodiscus axenaria.
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