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变异链球菌RgpAc基因敲除突变株的构建

Construction of RgpAc knock-out strain of S.mutans UA159
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摘要 目的:构建变异链球菌RgpAc基因敲除突变株,为进一步研究变异链球菌RgpAc基因功能做准备。方法:通过PCR扩增RgpAc基因上下游序列,分别插入到pFW5载体的2个多克隆酶切位点中,获得重组质粒。采用同源重组的方式将重组质粒转化到变异链球菌UA159中,获得RgpAc基因敲除突变株。结果:聚合酶链式反应(PCR)显示,突变株RgpAc基因已被载体完全替换。结论:成功构建变异链球菌RgpAc基因敲除突变株。 AIM: To construct a RgpAc knockout strain of Streptococcus mutans. METHODS: The up- stream and downstream sequences of RgpAc gene were amplified by polymerase chain reaction (PCR) , and were then inserted into 2 multiple clone enzyme cutting sites of pFW5 vector. The reconstructed plasmid was transformed into Streptococcus mutans UA159 by homologous recombination to get the RgpAc gene knock-out mutant. RESULTS: PCR and fluorescence quantitative PCR results showed that the RgpAc gene was replaced. CONCLUSION: Streptococcus mutans RgpAc knockout mutant strain was successfully constructed.
出处 《牙体牙髓牙周病学杂志》 CAS 北大核心 2013年第6期380-383,共4页 Chinese Journal of Conservative Dentistry
基金 国家自然科学基金项目(30672327 81100747)
关键词 变异链球菌 RgpAc pFW5载体 同源重组 streptococcus mutans RgpAc pFW5 vector homologous recombination
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