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中国对虾血蓝蛋白基因cDNA的克隆与序列分析
孙杰1, 王宝杰2, 孙姝娟2, 李晓华3, 姜珊2, 刘梅3, 蒋克勇3, 王雷3
1中国科学院海洋研究所,青岛 266071)中国科学院研究生院,北京 100039,山东省花生研究所,青岛 266100;2.中国科学院海洋研究所,青岛 266071,中国科学院研究生院,北京 100039;3.中国科学院海洋研究所,青岛 266071
摘要:
利用3’和5’RACE技术从中国对虾Fenneropenaeus chinensis肝胰腺中克隆了1个血蓝蛋白基因FCHc, FCHc基因cDNA全长为2 161bp。其中,开放阅读2 034bp,编码678个氨基酸,预测分子量为77.59 kDa。FCHc序列与凡纳滨对虾Litopenaeus vannamei血蓝蛋白同源性为82%,与日本囊虾Marsupenaeus japonicas同源性为85%。Real-timePCR实验结果表明,FCHc在肝胰腺中的相对表达量最高,在心脏和表皮中几乎不表达。该基因在鳗弧菌和对虾白斑综合征病毒(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,提示中国对虾FCHc基因在免疫反应中具有重要作用。
关键词:  中国对虾  血蓝蛋白  基因克隆  序列分析
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基金项目:国家自然科学基金青年基金项目(30600458)和国家高技术研究发展计划(863计划)项目(2006AA100311)共同资助
cDNA cloning and sequence analysis of hemocyanin in Fenneropenaeus chinensis
Abstract:
A hemocyanin gene (FCHc) was cloned from hepatopancreas of Chinese shrimp Fenneropenaeus chinensis by Rapid Amplification Complementary DNA Ends (RACE) method. The full length cDNA of FCHc consists of 2 161bp with a 2 034 bp Open Reading Frame (ORF),which encoded 678 amino acids,and the predicted molecular mass was 77.59kDa. It showed 82% and 85% homology with those of Litopenaeus vannamei and Marsupenaeus japonicas. The real-time PCR results showed that the expression level of FCHc in hepatopancreas was the highest among seven studied tissues.Real-time PCR analysis showed that the expression of FCHc was up-regulated distinctly in the hepatopanereas of shrimp when they were challenged by Vibrio anguillarum and white spot syndrome virus (WSSV). FCHc showed different expression profiles in the hepatopancreas during V. anguillarum or WSSV virus infection.The results implied that FCHc might play an important role in shrimp immune system.
Key words:  Fenneropenaeus chinensis  Hemocyanin  Gene cloning