耐辐射奇球菌pprM DNA结合域功能分析预测与基因敲除重组体的构建  

Bioinformatics analysis of pprM DNA binding domain and construction of gene knockout vector recombinant in deinococcus radiodurans

作　　者：杨杰[1] 苏泽红[2] 李斌元[2] 马云[2] 肖潇[1] 范美婷[1] 李伟[2] 王秋岩[2] 何淑雅[1,2] 

机构地区：[1]南华大学公共卫生学院,衡阳421001 [2]南华大学生物化学与分子生物学教研室,衡阳421001

出　　处：《辐射研究与辐射工艺学报》2013年第6期19-23,共5页Journal of Radiation Research and Radiation Processing

基　　金：国家自然科学基金项目(81272993)资助

摘　　要：通过生物信息学分析发现,pprM基因含有冷休克DNA结合域。利用Overlap PCR方法获得pprM基因DNA结合域缺失的基因片段pprMΔCSD后,酶切扩增的片段和自杀质粒空载体pk18mobsacB,并将目的基因片段与自杀质粒载体连接构建重组载体pk18mobsacB-pprMΔCSD,将重组子转化大肠杆菌JM109感受态,筛选出阳性克隆,酶切及测序鉴定。成功构建pprM DNA结合域基因敲除重组体,且生物信息学分析预测结果显示pprM DNA结合域与耐辐射奇球菌抗辐射功能相关。Bioinformatics analysis showed that pprM gene contained a Cold-shock DNA-binding domain. The pprMACSD was obtained by Overlap-PCR technology. And then it was connected with the suicide vector pklSmobsacB. Before ligation, amplified fragment and the empty vector were digested by the restriction enzyme EcoRI and BamHI. The ligation products were transformed into E.coil JM109. Positive clones were selected and followed by restriction enzyme digestion and sequencing analysis results. The pprM gene DNA binding domain knockout vector recombinants were successfully constructed. The bioinformatics analysis shows that pprM gene DNA binding domain is associated with the radiation resistance function in Deinococcus radiodurans.

关 键 词：耐辐射奇球菌 pprM基因 重叠延伸PCR 电离辐射 

分 类 号：Q78[生物学—分子生物学] TL71[核科学技术—辐射防护及环境保护]