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耐辐射球菌DNA修复开关基因pprI缺陷性突变和功能补偿性突变株的建立 被引量：4

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摘要 PprI是近来在耐辐射球菌Deinococcus,radiodurans中发现的一个极其重要的DNA修复开关基因.以已测序的野生型菌株R1为材料,运用PCR突变法将克隆具有自身GroEL启动子和卡那霉素抗性基因的DNA片段反向重组到pprI基因中去,首次获得了卡那霉素抗性的、pprI功能完全破坏的突变株YR1.辐射细胞生存率结果表明,YR1对辐射异常敏感.另外,将含有GroEL启动子和卡那霉素抗性基因的DNA片段重组到质粒pRADZ3中,获得了具有卡那霉素抗性的表达质粒pRADK;再将体外克隆的具有pprI完整基因和C-末端结构域截短的PprI蛋白基因片段分别重组到pRADK中.研究结果表明,含有pprI完整基因的表达质粒在YR1中能够正常表达,并完全恢复到野生型的极端抗性;而C-末端结构域截短的PprI蛋白基因片段虽能在YR1中正常表达,但对辐射异常敏感,不能恢复其极端抗性.上述pprI基因功能缺陷性和功能补偿性突变株的建立,为深入研究细胞体内PprI蛋白质的定位、结构域与功能的关系,以及原位研究PprI蛋白质的理化特性提供了十分有效的方法.
作者 高冠军 陆辉明 黄丽芬 华跃进
机构地区 浙江大学原子核农业科学研究所
出处 《科学通报》 EI CAS CSCD 北大核心 2005年第3期232-237,共6页 Chinese Science Bulletin
基金 国家重点基础研究发展计划(课题编号:2004CB719604) 国家自然科学基金(批准号:30330020) 国家杰出青年基金(批准号:30425038)资助项目.
关键词 性基因 DNA修复 正常 表达质粒 细胞 截短 首次 结构域 卡那霉素抗性 耐辐射球菌
分类号 Q78 [生物学—分子生物学]
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参考文献20

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二级参考文献23

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共引文献12

同被引文献91

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二级引证文献7

科学通报
2005年 第3期

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