多药耐药草绿色链球菌LuxS基因同源重组质粒的构建

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (10): 984-987.

多药耐药草绿色链球菌LuxS基因同源重组质粒的构建

陆笑¹,刘少娟¹,章锦才^2*,刘芹¹,林珊¹,彭湘明¹

1. 广州市红十字会医院口腔科广东广州 510220;
2. 南方医科大学附属口腔医院·广东省口腔医院广东广州 510280

收稿日期:2015-03-04 出版日期:2015-10-28 发布日期:2016-05-04
通讯作者: 章锦才,E-mail: zhwxluxiao@163.com
作者简介:陆笑(1975~ ),男,山东淄博市人,硕士,副主任医师,主要从事口腔科的临床治疗工作。
基金资助:
广州市卫生局基金资助项目(编号:2060402)

Construction of Streptococcus Viridans LuxS Gene Homologous Recombinant Plasmid

LU Xiao¹, LIU Shao-juan¹, ZHANG Jin-cai², LIU Qin¹, LIN Shan¹, PENG Xiang-ming¹

1. Guangzhou Red Cross Hospital, Guangzhou 510220, China;
2. Affiliated Stomatological Hospital of Southern Medical University, Guangdong Provincial Stomatological Hospital, Guangzhou 510280, China

Received:2015-03-04 Online:2015-10-28 Published:2016-05-04

摘要/Abstract

摘要： 目的:构建一个含有红霉素抗性基因和多药耐药草绿色链球菌LuxS基因上下游区同源序列的重组质粒,为后续进行LuxS基因敲除实验做准备。方法:设计合成引物,分别以质粒PMG36E、草绿色链球菌DNA为模板,应用聚合酶链反应(PCR)扩增得到红霉素抗性基因DNA片断和LuxS基因上下游序列,经双酶切反应插入pUC19质粒的多克隆酶切位点中,转化大肠杆菌的感受态中,氨苄青霉和红霉素培养基筛选。结果:红霉素抗性基因和LuxS基因两侧同源序列成功连入到Puc19质粒相应酶切位点,PCR凝胶电泳、测序结果正确。结论:成功构建多药耐药草绿色链球菌LuxS基因敲除同源重组质粒,为进一步构建LuxS突变株打下基础。

关键词: 草绿色链球菌, LuxS基因, pUC19质粒, PMG36E质粒

Abstract: Objective: To construct a recombination plasmid containing erythromycin resistance gene and the upstream and downstream homologous sequence of LuxS gene of streptococcus viridans (S. viridans) with multidrug resistance. Methods: Erythromycin resistance gene and the upstream and downstream homologous sequence of LuxS were cloned respectively by using plasmid PMG36E and DNA of S. viridans template. Then the genes were ligated into multiple cloning site (MCS) of vector pUC19 and transformed into competent E. coli. Finally, the transformants which were resistant to erythromycin and ampicillin were selected. Results: Erythromycin resistance gene and the upstream and downstream homologous sequence of LuxS were successfully ligated into enzyme digestion site of vector pUC19 accurately. The PCR agarose gel electrophoretic analysis and the sequencing results were correct. Conclusion: The S. viridans recombinant plasmid with LuxS geneknock-out was constructed and could be used for constructing S. viridans LuxS mutans in future.

Key words: Streptococcus viridans, LuxS gene, Vector pUC19, Plasmid, PMG36E

中图分类号:

R780.1

陆笑,刘少娟,章锦才,刘芹,林珊,彭湘明. 多药耐药草绿色链球菌LuxS基因同源重组质粒的构建[J]. 口腔医学研究, 2015, 31(10): 984-987.

LU Xiao, LIU Shao-juan, ZHANG Jin-cai, LIU Qin, LIN Shan, PENG Xiang-ming. Construction of Streptococcus Viridans LuxS Gene Homologous Recombinant Plasmid[J]. Journal of Oral Science Research, 2015, 31(10): 984-987.

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