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- 2018
变形链球菌耐氟菌株gtfB基因失活菌株的构建
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Abstract:
摘要 目的:构建变形链球菌(S.mutans)耐氟菌株UA159-FR中gtfB基因失活株,为研究耐氟菌株gtfB基因的功能奠定基础。方法:培养UA159-FR并以其为模板扩增gtfB基因上游、下游同源臂片段,以质粒pEGFP-N1为模板扩增kan基因;通过重叠延伸聚合酶链反应法(overlap extension polymerase chain reaction,OE-PCR)获取上述3个片段的同源重组片段;与pEASY-Blunt Cloning Vector连接形成重组质粒后,进行PCR鉴定和测序鉴定;将重组片段电转化入UA159-FR感受态细胞中得到gtfB基因失活菌株,并进行PCR鉴定。结果:经PCR鉴定和测序鉴定,含有gtfB基因重组片段的重组质粒构建成功;经PCR鉴定,UA159-FR的gtfB基因失活株构建成功。结论:成功构建了含有gtfB基因重组片段的重组质粒和S.mutans耐氟菌株UA159-FR的gtfB基因失活株,可用于gtfB基因功能的研究
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