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中国明对虾caspase 基因的克隆与表达分析
宋光年1,2, 金松君1, 张继泉1, 相建海1
1.中国科学院海洋研究所;2.中国科学院研究生院
摘要:
利用细胞凋亡来清除多细胞器官受损伤或有害的细胞, 为动物发育过程中保持机体平衡起到关键作用。为了研究细胞凋亡在甲壳类动物应对病毒感染后所起到的作用, 作者利用同源克隆技术获得了中国明对虾(Fenneropenaeus chinensis) caspase 基因(FcCasp)的cDNA 序列, 并分析了该基因在对虾受到白斑综合症病毒(white spot syndrome virus,简称WSSV)刺激后的表达变化规律。结果表明, 该基因的cDNA 含有954 个核苷酸, 编码317 个氨基酸, 具有caspase 家族典型的QACRG 五肽蛋白酶活性位点(190-194 氨基酸)。对注射了WSSV 的中国明对虾FcCasp 的表达进行分析, 在5 个时间点取其肝胰脏样本, 经real-time PCR 检测发现, WSSV 刺激3 h 后, 对虾肝胰脏中FcCasp 的表达呈现显著性上调,说明FcCasp 的表达与WSSV 感染密切相关, 提示FcCasp 基因与细胞凋亡相关。
关键词:  中国明对虾(Fenneropenaeus chinensis)  基因克隆  Caspase  WSSV 攻毒  实时定量PCR 分析
DOI:
分类号:
基金项目:国家973 资助项目(2006CB101804); 国家863 计划项目(2006AA10A401); 国家自然基金项目(30970422)
Cloning and expression of a caspase gene in Chinese shrimp Fenneropenaeus chinensis
Abstract:
Apoptosis eliminates unwanted cells, from an organism including damaged and virus-infected cells, being a fundamental cellular process that plays a critical role in normal development and tissue homeostasis. Here we studied a caspase homolog from Chinese shrimp Fenneropenaeus chinensis, designated FcCasp. An open reading frame of 954 nucleotides encoding 317 amino acids (aa) contained a conserved motif, QACRG, of known caspases-3, and was located at aa 190–194. FcCasp expressions were measured in normal shrimp and in shrimp infected by white spot syndrome virus (WSSV) at five time-points post-injection (p.i.) with real-time PCR, revealng different expression profiles of FcCasp gene in hepatopancreas of shrimps after they were challenged by WSSV. The transcription of FcCasp in hepatopancreas showed up-regulation 12 h post-challenge, indicating the expression of FcCasp was closely related to WSSV challenge. Therefore, FcCasp might be involved in the apoptosis.
Key words:  Fenneropenaeus chinensis  caspase gene cloning  WSSV challenge  real-time PCR
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