耐辐射奇球菌RecR和SSB蛋白的克隆表达及抗紫外功能
Chinese Journal of Appplied Environmental Biology(2010)
Abstract
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.图2表2参14
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Key words
recR,ssb,gene clone,Deinococcus radiodurans,UV resistance
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