| 摘要: |
| 本文在对中国明对虾转录组数据进行生物信息学分析的基础上, 克隆获得了其Akt/PKB基因(FcAkt)的开放阅读框(ORF)的cDNA序列, 分析了对虾FcAkt基因的结构特征, 并利用实时定量RT-PCR技术分析了FcAkt基因在鳗弧菌、溶壁微球菌和白斑综合征病毒(WSSV)注射后的转录表达变化。结果表明, 中国明对虾FcAkt基因的ORF全长为1536bp, 编码511个氨基酸, 预测蛋白分子量为58.9kDa, 理论等电点pI为5.60。同源比对显示该基因的氨基酸序列在不同物种之间具有较高的保守性。进化分析发现FcAkt与地蟹、果蝇、埃及伊蚊、家蚕等节肢动物的蛋白激酶B(Akt/PKB)亲缘关系较近。FcAkt的转录表达在不同细菌或WSSV刺激后具有明显的变化, 提示对虾FcAkt基因可能在对虾的免疫防御中发挥了重要作用。 |
| 关键词: 中国明对虾, Akt/PKB, 基因克隆, 表达分析 |
| DOI：10.11693/hyhz201303011011 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划(973)项目, 2012CB114403; 国家虾产业体系项目, CARS-47 号 |
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| EXPRESSION AND FUNCTION ANALYSIS OF Akt/PKB IN PATHOGENS-CHALLENGED CHINESE SHRIMP FENNEROPENAEUS CHINENSIS |
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CHI Yan-Hong1,2, LI Shi-Hao1,2, WANG Dong-Dong1,2, SUN Yu-Miao1, LI Fu-Hua1, XIANG Jian-Hai1
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1.Institute of Oceanology, Chinese Academy of Sciences;2.University of Chinese Academy of Sciences
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| Abstract: |
| Akt/PKB is a serine/threonine protein kinase that plays a key role in multiple cellular processes. Based on the bioinformatics analysis of transcriptome data, an Akt/PKB gene (FcAkt) was obtained from the Chinese shrimp Fenneropenaeus chinensis and the protein structures of FcAkt was analyzed. Real-time PCR was applied to study the transcription profiles of FcAkt after Gram-positive (G+) bacteria (Micrococcus lysodeikticus), Gram-negative (G-) bacteria (Vibrio anguillarium) or WSSV injection in shrimp. The open reading frame (ORF) of FcAkt consists of 1536bp encoding 511aa. The predicted molecular weight (MW) of the deduced protein of FcAkt was 58.92kDa, and its theoretical pI was 5.60. Homology analysis showed that the amino acid sequence of Akt had high conservation among different species, and phylogenetic analysis showed that FcAkt and Akt from other arthropods had close genetic relationship. The transcription of FcAkt showed apparent changes after bacteria or WSSV challenge. The present data provide clues that FcAkt might play potential important roles in the innate immunity of shrimp. |
| Key words: Fenneropenaeus chinensis Akt/PKB Gene cloning Expression analysis |
