4个不同地理群体中华鳖pomc基因PCR-RFLP分析
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中央级公益性科研院所基本科研业务费专项(2011JBFA19)


PCR-RFLP analysis on pomc gene in four populations of Pelodoiscus sinensis
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    摘要:

    为探讨不同地理群中华鳖的pomc基因的多态性,建立区分中华鳖不同地理群的分子遗传标记,采用PCR-RFLP的方法扩增得到4个不同地理群(太湖鳖群体、沙鳖群体、台湾鳖群体和黄河鳖群体)中华鳖的pomc基因,比较4个地理群中华鳖的pomc基因的同源性,构建遗传进化树,对其亲缘关系进行分析,并用AccⅡ、BscBⅠ、AsuⅠ和TscⅠ4种限制性内切酶对pomc基因进行酶切分析。结果显示: 4个不同地理群的中华鳖均能扩增出786 bp的pomc基因。亲缘关系分析表明:在遗传上台湾鳖和沙鳖亲缘关系较近,而太湖鳖与黄河鳖亲缘关系较近。用酶切分析pomc基因扩增产物,结果表明:用AccⅡ内切酶对4种中华鳖pomc基因进行酶切,其中沙鳖切出303 bp和483 bp 2个条带,而其他3个种属的中华鳖只有1个786 bp的条带。用BscBⅠ酶对4种中华鳖pomc基因进行酶切,其中台湾鳖和沙鳖切出351 bp和435 bp 2个条带,而其他2个种属的中华鳖只有1个786 bp的条带。用AsuⅠ酶对4种中华鳖pomc基因进行酶切,其中台湾鳖和黄河鳖切出349 bp和437 bp 2个条带,而其他2个种属的中华鳖只有1个786 bp的条带。用TscⅠ酶对中华鳖pomc基因进行酶切,其中黄河鳖和太湖鳖切出349 bp和437 bp 2个条带,而其他2个种属的中华鳖只有1个786 bp的条带。这4个限制性内切酶的联合使用分析,可使这4个地理群的中华鳖在分子水平都得到明确的鉴定。从这4个地理群的中华鳖pomc基因核苷酸序列的显著差异和酶切位点的变化,进一步证明pomc基因可以作为区分这4个不同地理群中华鳖的分子遗传标记。

    Abstract:

    To establish a molecular genetic marker for identifying different populations of Pelodoiscus sinensis,pomc gene was amplified by PCR and identified by RFLP using four restriction endonucleases including AccⅡ,BscBⅠ,AsuⅠ and TscⅠ.At the same time homology were compared and phylogenetic trees were constructed.The results showed that a 786 bp fragment was amplified from four P.sinensis populations.The relationship between Taiwan-introduced and Pelodiscus axenaria populations was closer and the relationship between Taihu Lake and Huanghe River population was closer.For AccⅡ digestion,the P.axenaria population carried two bands of 303 bp and 483 bp,while the other three populations carried only one band of 786 bp; for BscBⅠ digestion,Taiwan-introduced and P.axenaria population carried two bands of 351 bp and 435 bp,while the other two populations carried only one band of 786 bp; for AsuⅠ digestion,Taiwan-introduced and Huanghe River population carried two bands of 349 bp and 437 bp,while the other two populations carried only one band of 786 bp; for TscⅠ digestion,Taihu Lake and Huanghe River populations carried two bands of 349 bp and 437 bp,while the other two populations carried only one band of 786 bp.Combined application of the four restriction endonuclease could clearly identify the different population of P.sinensis at the molecular level.All results suggested that the pomc gene could be used as a molecular genetic marker for identifying different population of P.sinensis.

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张林,吴涛,何力.4个不同地理群体中华鳖pomc基因PCR-RFLP分析[J].华中农业大学学报,2013,32(5):106-111

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  • 收稿日期:2013-01-02
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