胞内分枝杆菌HSP65基因克隆、分析及其原核表达的研究  

Cloning of Mycobacterium intracellulare HSP 65 and its analysis and prokaryotic expression

作　　者：葛淑敏[1,2] 王诚[1] 王准[1] 钱爱东[2] 

机构地区：[1]长春理工大学生命科学技术学院,吉林长春130022 [2]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《现代预防医学》2015年第3期515-518,532,共5页Modern Preventive Medicine

基　　金：国家自然基金项目(31272566);吉林省科技发展计划项目(20120235);省级创新训练项目(2014S042)

摘　　要：目的克隆胞内分枝杆菌HSP 65基因,对其进行生物信息学分析与原核表达。方法应用巢式PCR克隆胞内分枝杆菌HSP 65基因,利用在线分析软件对其进行生物信息学分析。构建原核表达载体p ET 15b-HSP 65并进行诱导表达。结果胞内分枝杆菌HSP 65基因完整ORF基因全长1 626 bp,编码541个氨基酸。HSP 65蛋白为酸性、亲水性蛋白质,亚细胞定位主要存在于细胞质,蛋白二级结构以α-螺旋、无规则卷曲及β-折叠为主。使用同源建模法预测了HSP 65基因编码蛋白三维立体结构。试验构建p ET 15b-HSP 65原核表达载体并成功表达。结论成功克隆胞内分枝杆菌HSP 65基因,并对HSP 65蛋白进行了生物信息学分析及原核表达。Objective To study cloned prokaryotic expression of the Mycobacterium intracellulare HSP 65 gene and analyze the biological information. Methods The Mycobacterium intracellulare HSP 65 gene was amplified by nested PCR and the biological information analysis was performed. At the same time, the prokaryotic expression vector p ET 15b-HSP 65 was constructed and inducted expression in E. coli BL21 p Lys S. Results The Mycobacterium intracellulare HSP 65 has been cloned. Its ORF consisted of 1626 nucleotides, encoding 541 amino acids. Mycobacterium intracellulare HSP 65 is an acid and hydrophilic protein. Most of HSP 65 exist in the cytoplasmic and the main structures of the protein are α-helix, random coil and β-pleate. At the same time,the tertiary structure was also predicted by homology modeling. In this experiment, the p ET 15b-HSP 65 as a prokaryotic expression vector was successfully constructed, and it got a good expression. Conclusion The Mycobacterium intracellulare HSP 65 has been successfully cloned, and the p ET 15b-HSP 65 as a Prokaryotic expression vector got a good expression.

关 键 词：胞内分枝杆菌 HSP 65 克隆 生物信息学分析 原核表达 

分 类 号：R113[医药卫生—公共卫生与预防医学]