中国明对虾酚氧化酶原基因cDNA的克隆与表达特征

The full length cDNA cloning and expression profile of prophenoloxidase of Fenneropenaeus chinensis

  • 摘要: 利用3′和5′RACE技术从中国明对虾血细胞中克隆了一个酚氧化酶原基因FCproPO, FCproPO基因cDNA全长为2 311 bp,其中开放阅读框 2 061 bp,编码687个氨基酸,预测分子量为78.71 ku。推测的序列与凡纳滨对虾同源性为84%,与日本囊对虾同源性为71%。RT-PCR实验结果表明,FCproPO在血细胞中的相对表达量最高,在心脏和精巢中几乎不表达。序列分析发现FCproPO含有两个保守的铜离子结合位点;进化分析发现FCproPO与斑节对虾、日本囊对虾、凡纳滨对虾等海水虾的酚氧化酶原亲缘关系最近。该基因在被鳗弧菌和对虾白斑病毒(WSSV)感染后的对虾肝胰腺中的表达量显著增加,并具有不同的时空表达趋势,提示中国明对虾FCproPO基因在免疫反应中具有重要作用。

     

    Abstract: Chinese shrimp (Fenneropenaeus chinensis) is one of the most economically valuable and widely cultured species in China. During the last decade its development has been seriously affected on a regular basis by the outbreak of viral and bacterial diseases. To analyze the immune defense mechanisms of shrimp F. chinensis, a prophenoloxidase gene (FCproPO) was cloned from haemocytes of Chinese shrimp by Rapid Amplification Complementary DNA Ends (RACE) method. The full length cDNA of FCproPO consisted of 2 311 bp with a 2 061 bp Open Reading Frame(ORF),which encoded 687 amino acids,and the predicted molecular mass was 78.71 ku. It showed 84% and 71% homology with those of Litopenaeus vannamei and Marsupenaeus japonicas. The Realtime-PCR results showed that the expression level of FCproPO in haemoeytes was highest among seven studied tissues.Sequence analysis showed FCproPO contained two conserved copper-binding sites. Phylogenetic analysis revealed that FCproPO and prophenoloxidase from L. vannamei, M. japonicus and P. monodon were in the same phylogenetic branches. Realtime-PCR analysis showed that the expression of FCproPO was up-regulated distinctly in the hepatopanereas of shrimp when they were challenged by Vibrio anguillarum and white spot syndrome virus (WSSV). FCproPO showed different expression profiles in the hepatopancreas during V. anguillarum or WSSV virus infection.The result implied that FCproPO might play an important role in the shrimp immune system.

     

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