上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (7): 875-.doi: 10.3969/j.issn.1674-8115.2011.07.002

• 论著（基础研究） • 上一篇    下一篇

Rv1272和Rv1273与结核分枝杆菌耐药相关性研究

程旭红1,2, 孙 青1, 姚玉峰2, 周爱萍2,3, 赵英伟1   

  1. 1．苏州大学 基础医学与生物科学学院, 苏州 215123； 2．上海交通大学 基础医学院病原生物学教研室, 上海 200025； 3．西藏民族学院, 咸阳 712082
  • 出版日期:2011-07-28 发布日期:2011-07-27
  • 通讯作者: 赵英伟, 电子信箱: zhaoyingwei1993@sina.com；周爱萍, 电子信箱: abby_aiping@126.com。
  • 作者简介:程旭红(1983—), 女, 硕士生；电子信箱: chengxuhong0529@163.com。
  • 基金资助:

    国家重点基础研究发展计划(“973”计划) (2009CB522605)资助项目

Association of Mycobacterium tuberculosis Rv1272 and Rv1273 with drug resistance

CHENG Xu-hong1,2, SUN Qing1, YAO Yu-feng2, ZHOU Ai-ping2,3, ZHAO Ying-wei1   

  1. 1.School of Biology and Basic Medical Sciences, Suzhou University, Suzhou 215123, China；2.Department of Medical Microbiology and Parasitology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；3.Tibet University for Nationalities, Xianyang 712082, China
  • Online:2011-07-28 Published:2011-07-27
  • Supported by:

    National Key Basic Research Development Program, “973” Program, 2009CB522605

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摘要：

目的 研究结核分枝杆菌Rv1272和Rv1273基因编码蛋白与药物外排的关系。方法 利用生物信息学方法分析Rv1272和Rv1273基因及其编码蛋白的结构，采用PCR技术扩增该两个基因，并与pMF406质粒连接构建重组质粒，测序正确后，电穿孔法将重组质粒转化耻垢分枝杆菌mc2155。采用Western blotting对两个目的蛋白进行亚细胞定位；试管法测定重组菌株对常用的9种抗菌药物的最低抑菌浓度（MIC），以转化pMF406空质粒的耻垢分枝杆菌作为对照菌株。结果 经测序验证重组质粒构建成功，Western blotting结果显示Rv1272和Rv1273为膜蛋白。试管法测定结果显示：含有目的基因的重组菌株对9种抗菌药物的MIC与对照菌株比较，差异无统计学意义（P>0.05）。结论 Rv1272和Rv1273蛋白为膜蛋白，未发现其与结核菌耐药之间的直接关系。

关键词: 结核分枝杆菌, Rv1272, Rv1273, 膜蛋白, 最低抑菌浓度

Abstract:

Objective To investigate the association of two proteins coded by Mycobacterium tuberculosis Rv1272 and Rv1273 with drug efflux. Methods The structures of Rv1272 gene, Rv1273 gene and proteins coded by them were analyzed by bioinformatics, and these two genes were amplified by PCR and cloned into expression vector pMF406 to generate the recombinant plasmids. After verification by sequence analysis, the recombinant plasmids were transformed into M.Smegmatis mc2155 by electroporation. The subcellular localization of these two proteins was probed by Western blotting, and the minimal inhibitory concentrations (MIC) of nine commonly used antibiotics drugs were determined by test tube method, with M. Smegmatis as control strain. Results It was verified by sequence analysis that the recombinant plasmids were successfully constructed, and it was identified by Western blotting that Rv1272 and Rv1273 were membrane proteins. It was revealed by test tube method that there was no significant difference in MIC of nine commonly used antituberculosis drugs between recombinant strains and control strain (P>0.05). Conclusion Rv1272 and Rv1273 are membrane proteins, and their relationship with drug resistance to Mycobacterium tuberculosis is uncertain.

Key words: Mycobacterium tuberculosis, Rv1272, Rv1273, membrane protein, minimal inhibitory concentration