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为研究凡纳滨对虾适应低温的分子机理,实验对从低温处理凡纳滨对虾抑制性消减文库中筛选出的一个360 bp EST序列进行了研究。首先,同源比对显示该EST片段与其他物种的ANT2基因高度同源,命名为凡纳滨对虾ANT2基因(LVANT2);其次,通过构建凡纳滨对虾肝胰腺全长cDNA文库,PCR扩增获得LVANT2基因的全长cDNA1540 bp,其中包括1011 bp的完整开放阅读框,编码336个氨基酸残基。然后,对基因进行了不同组织和低温处理的表达谱分析:(1)组织表达谱的结果显示,该基因在凡纳滨对虾肌肉组织中表达量最高;(2)在不同低温处理下的表达结果显示,该基因在15℃处理下基因表达量发生显著变化,13℃开始呈下调表达,11℃时表达量又升高;13℃低温处理不同时间发现该基因在12h内表达量发生显著变化,48h后表达量最高。LVANT2基因的低温诱导表达模式说明其可能在凡纳滨对虾低温适应中发挥作用。
In order to study the molecular mechanism of low temperature adaptation in Litopenaeus vannamei, a 360 bp EST sequence screened from cryogenic suppression library of Litopenaeus vannamei was studied. First of all, homologous alignment showed that the EST fragment was highly homologous to the ANT2 gene of other species and was named as the ANT2 gene of V. vannamei (LVANT2). Secondly, the full-length hepatopancreas cDNA library was constructed by PCR amplification The full-length cDNA of LVANT2 gene was 1540 bp, including a complete open reading frame of 1011 bp, which encoded 336 amino acid residues. Then, the gene expression profile of different tissues and hypothermia were analyzed: (1) The result of tissue expression profile showed that the gene was expressed most in Muscle tissue of Litopenaeus vannamei; (2) The expression under different hypothermia The results showed that the gene expression at 15 ℃ had a significant change, which was down-regulated at 13 ℃ and increased at 11 ℃. The expression level of the gene at 12 ℃ was significantly changed at 12 ℃ , 48h after the highest expression. The low temperature induced expression pattern of LVANT2 gene indicates that it may play a role in low temperature adaptation of Litopenaeus vannamei.