耐辐射奇球菌RecR和SSB蛋白的克隆表达及抗紫外功能  

Cloning Expression and UVC Resistant Function of RecR and SSB Proteins from Deinococcus radiodurans

作　　者：常晓松[1,2] 杨澜[1] 付文娟[1] 赵清[1] 胡冉[1] 陈济安[1] 舒为群[1] 

机构地区：[1]重庆第三军医大学环境卫生学教研室,重庆400038 [2]广州军区武汉总医院检验科,武汉430070

出　　处：《应用与环境生物学报》2010年第3期328-331,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(Nos.30670635;50804049);全军医药卫生科研基金项目(No.06M0199);重庆市自然科学基金项目(No.CSTC2007BB5004)资助~~

摘　　要：RecR和SSB蛋白是耐辐射奇球菌(Deinococcus radiodurans)同源重组修复RecFOR途径的关键蛋白.以耐辐射奇球菌基因组为模板,PCR扩增recR和ssb基因片段,以质粒pET32a(+)为载体,构建重组质粒pET32a(+)-recR和pET32a(+)-ssb,并转化到Escherichia coli BL21(DE3)中,IPTG诱导蛋白表达,表达产物采用SDS-PAGE鉴定,同时,应用平板菌落计数法检测紫外辐照前后各组细菌生存率变化,研究RecR和SSB蛋白在E.coli BL21(DE3)抗紫外线辐射中的作用.结果表明,RecR和SSB能够克隆至pET32a(+)载体并在E.coli BL21(DE3)中诱导表达.紫外辐照生存率实验结果提示,RecR和SSB蛋白能增强E.coli BL21(DE3)对紫外线辐射的抵抗能力.RecR和SSB的成功表达及紫外抗性特点将为耐辐射奇球菌超强耐辐射机制的研究提供实验基础.To construct the expression recombinants of RecR and SSB from Deinococcus radiodurans,express these target proteins in Escherichia coli BL21（DE3） and analyze UVC resistant abilityies of RecR and SSB in E.coli,recR and ssb genes were amplified by PCR from genomic DNA of D.radiodurans and inserted into expression plasmid vector pET32a（＋） to construct pET32a（＋）-recR and pET32a（＋）-ssb.The recombinant plasmids were then transformed into E.coli BL21（DE3） and the recombinant proteins were expressed by induction with isopropyl-β-D-thiogalactopyranoside（IPTG） and analyzed with SDS-PAGE.Compared to the survival rate of bacteria in each group before and after UVC irradiation,the recombinant plasmids pET32a（＋）-recR and pET32a（＋）-ssb were obtained and these recombinant proteins could be highly expressed in E.coli BL21（DE3）.The results indicated that RecR and SSB from D.radiodurans could be expressed in E.coli BL21 successfully and could increase UVC resistance of E.coli.This study provides basic data for further studies on and future applications of the recombinants RecR and SSB.

关 键 词：recR SSB 基因克隆 耐辐射奇球菌 紫外抗性 

分 类 号：Q78Q939.105[生物学—分子生物学]