耐辐射奇球菌DR_2327和DR_2328基因克隆及功能分析

通过生物信息学分析耐辐射奇球菌DR_2327和DR_2328基因及编码蛋白的基本性质,利用PCR方法克隆DR_2327和DR_2328的全基因,连接至p GEM-T载体上,转化至大肠杆菌JM109中,并进行双酶切鉴定。生物信息学分析结果表明,DR_2327和DR_2328基因位于同一操纵子中,且DR_2328蛋白具有组氨酸激酶活性,DR_2327蛋白具有反应调节子功能,提示两者很可能组成一对双组分系统。体外实验表明,DR_2327和DR_2328基因的外源表达在一定程度上提高了大肠杆菌对紫外辐照和H2O2的耐受能力。

Cloning,and functional analysis ofDR_2327 andDR_2328 inDeinococcus radiodurans

The main characteristics ofDR_2327and DR_2328 inDeinococcus radiodurans were analyzeed by bioinformatics. Full-length gene ofDR_2327and DR_2328 was amplified by PCR, and constructed in pGEM-T vector. Transformed recombinant vector intoE.coli JM109 and followed by restriction enzyme digestion.DR_2327 andDR_2328 gene was cloned successfully. Bioinformatic analysis results showed thatDR_2327 andDR_2328 gene located in the same operon, and DR_2328 protein with histidine kinase activity, DR_2327 protein with response regulator activity. It is suggested thatDR_2327 andDR_2328 constitute a two-component system inDeinococcus radiodurans.In vitro experiments showed thatDR_2327 andDR_2328could enhance tolerance to UV radiation and H2O2 ofE.coli.