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陈璇;刘海霞;彭显;邹玲
华西口腔医学杂志, 2017, Volume: 35, Issue: 6Journal Article
目的利用框内缺失法,通过IFDC2基因盒及融合聚合酶链反应(PCR)、同源重组技术构建变异链球菌UA159菌株srtA基因缺失株。方法PCR扩增变异链球菌UA159菌株srtA基因上下游同源片段及IFDC2基因盒,将这些片段连接、转化入UA159,替换srtA基因同源片段,筛选、鉴定红霉素抗性克隆;PCR扩增、连接UA159srtA基因上下游同源片段,将连接片段转化人前述红霉素抗性克隆中替换IFDC2同源片段,筛选、鉴定抗苯丙氨酸类似物p-chloro-phenylalanine(p-C1-Phe)克隆。结果PCR、琼脂糖凝胶电泳及DNA测序结果均证明srtA基因编码区被完全删除,上下游片段无缝连接,成功构建UA159 srtA基因缺失株。结论本研究成功构建了无标记的变异链球菌UA159srtA基因缺失株,为进一步研究该基因在生物膜中的作用及其调控机制奠定了基础。
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