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凡纳滨对虾CTSL基因PCR-SSCP多态性与生长性状的关联分析及其mRNA的差异表达

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Keywords: 凡纳滨对虾,组织蛋白酶L基因,PCR-SSCP,关联分析,差异表达

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Abstract:

利用PCR-SSCP技术对379尾凡纳滨对虾组织蛋白酶CathepsinL(CTSL)基因多态性进行检测,采用最小二乘法对多态性与生长性状的相关性进行分析;利用Real-timequantitativePCR检测CTSL基因mRNA的差异表达情况。结果表明:引物C6(F/R)扩增片段具有多态性,其扩增产物具有CC、CT和TT3种基因型,TT基因型与CC基因型相比在第6外显子92bp处发生了C→T的突变,为沉默突变。最小二乘法分析结果表明,在体重、体长、第一腹节长和尾节长这4个指标上CC和CT基因型显著高于突变型TT(p<0.05),而在头胸甲长和头胸甲宽2个指标上,CC基因型显著高于CT和TT基因型(p<0.05)。CTSL基因mRNA在凡纳滨对虾各组织、不同性别个体肌肉组织、极端体重个体肌肉组织中相对表达量大小顺序分别为:肝胰脏>胃>眼柄>心脏>消化腺;雌虾>雄虾;极大个体>极小个体。结论:CTSL基因多态性对凡纳滨对虾生长性状有显著影响,可以作为影响该虾生长性状的候选基因,为该虾的选育提供分子遗传标记;CTSL基因mRNA的分布情况表明此基因可能正向调控对虾生长。

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