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

作　　者：孙杰[1,2,3] 王宝杰[1,2] 李晓华[1] 孙姝娟[1,2] 刘梅[1] 蒋克勇[1] 王雷[1] 

机构地区：[1]中国科学院海洋研究所,山东青岛266071 [2]中国科学院研究生院,北京100039 [3]山东省花生研究所,山东青岛266100

出　　处：《水产学报》2010年第1期56-66,共11页Journal of Fisheries of China

基　　金：国家"八六三"高技术研究发展计划(2006AA100311);国家自然科学基金项目(30600458)

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2311bp,其中开放阅读框2061bp,编码687个氨基酸,预测分子量为78.71ku。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。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 E anguillarum or WSSV virus infection. The result implied that FCproPO might play an important role in the shrimp immune system.

关 键 词：中国明对虾 酚氧化酶原 基因克隆 表达 

分 类 号：Q785[生物学—分子生物学] S917[农业科学—水产科学]