溶藻弧菌热休克蛋白70基因的克隆及序列分析  

Gene cloning and sequence analysis of HSP70 from Vibrio alginolyticus

作　　者：万文菊[1] 于爱莲[1] 刘含亮[2] 李庆雷[2] 张忠[1] 王纪亭[2] 杨春贵[1] 宋光乐[1] 

机构地区：[1]泰山医学院基础医学部,山东泰安271000 [2]山东农业大学动物科技学院,山东泰安271000

出　　处：《中国病原生物学杂志》2010年第7期498-501,共4页Journal of Pathogen Biology

基　　金：山东省优秀中青年科学家奖励基金项目(No.2007BS02017);泰山医学院博士启动金项目

摘　　要：目的通过测定溶藻弧菌热休克蛋白70(HSP70)基因序列,结合GenBank登录的其他弧菌HSP70核苷酸序列,分析溶藻弧菌HSP70特点以及与其他弧菌HSP70之间的同源性。方法根据GenBank中同类蛋白序列设计特异PCR引物,首次从溶藻弧菌中扩增热休克蛋白70ORF全长基因片段,并对该片段进行克隆、测序和分析。结果扩增的溶藻弧菌HSP70ORF全长基因片段长1914bp,与已登录的其他弧菌的同源性在90.4%～96.2%之间,包含完整的HSP70ORF,编码637个氨基酸。与GenBank中其他弧菌HSP70的氨基酸序列比较,溶藻弧菌HSP70含有Dnak特征基序DLGTT-S-V(8-16残基);整个分子的保守性N端最高,到C端保守性降低。结论溶藻弧菌与哈维弧菌(Vibrioharveyi)热休克蛋白70有较高同源性,该热休克蛋白70属于Dnak类型。Objective The objective of this study was to analyze the gene sequence of heat shock protein 70 of Vibrio alginolyticus and compare the gene sequence of heat shock protein 70 among vibrio stains reported in GenBank.MethodsA pair of primers was designed according to the nucleotide sequence of the heat shock protein 70 gene of vibrio reported in GenBank.With the specific primers,target fragments were amplified from Vibrio alginolyticus genomic DNA by PCR.The fragments were cloned and sequenced.Results Results indicated that the gene of HSP70 in Vibrio alginolyticus which shared 90.4%-96.2% homology of nucleotides with other vibrio stains,included an ORF of 1 914 bp and encoded a polypeptide of 637 amino acids that contained the characteristic sequence of Dnak （DLGTT-S-V）.The N-terminal amino acid sequence of the entire molecule was more conserved than the C terminal.Conclusion Results showed that the HSP70 isolated from Vibrio alginolyticus was a kind of Dnak that had a high homology of nucleotides with Vibrio harveyi.

关 键 词：热休克蛋白70 溶藻弧菌 基因克隆 

分 类 号：Q785[生物学—分子生物学]