冰岛硫化叶菌xpb基因缺失突变体的构建及遗传学分析
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国家自然科学基金项目(31100050,31128011)

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    摘要:

    为在体内研究古菌核苷酸切除修复(nucleotide excision repair,NER)机制,利用基因敲除的方法,对泉古菌核苷酸切除修复机制进行研究,构建了冰岛硫化叶菌编码真核生物NER解旋酶XPB同源蛋白的基因xpb1和xpb2单缺失突变体,从而在体内分析xpb1和xpb2基因的功能。结果表明:基因xpb1或xpb2不是冰岛硫化叶菌存活的必需基因。表型分析发现,xpb1和xpb2基因单缺失突变体相较野生型菌株REY15A,对DNA损伤试剂4-NQO 4-硝基喹啉-1-氧化物(4-nitroquinoline 1-oxide,4-NQO)分别表现出轻微敏感性和不敏感性,暗示NER解旋酶功能在冰岛硫化叶菌体内存在多重冗余。

    Abstract:

    Genome analyses suggested that archaea could possess a eukaryotictype nucleotide excision repair (NER) pathway because they encode homologs of several eukaryotic NER proteins.To investigate the archaeal nucleotide excision repair pathway in vivo,the genes encoding the potential NER helicases in the genome of the hyperthermophilic crenarchaeon Sulfolobus islandicus were deleted via a gene knockout strategy and deletion mutants of the xpb1 and xpb2 genes were isolated,indicating that they were not essential to the survival of S. islandicus.Phenotypic analysis of deletion mutants showed that the xpb1 mutant was only slightly sensitive to 4NQO,as compared with the wildtype strain REY15A,while the xpb2 mutant showed no sensitivity to 4NQO.It is the first genetic study of the crenarchaeon NER pathway.

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李素明,张昌毅,彭 楠,梁运祥,佘群新.冰岛硫化叶菌xpb基因缺失突变体的构建及遗传学分析[J].华中农业大学学报,2012,31(1):28-33

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  • 收稿日期:2011-04-03
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