Wang Qingzhong, Yue Jun, Zhang Lu, Xu Ying, Chen Jiazhen, Zhang Min, Zhu Bingdong, Wang Hongyan, Wang Honghai
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University and Shanghai Pneumology Hospital, Shanghai, PRC.
J Proteome Res. 2007 Dec;6(12):4564-71. doi: 10.1021/pr070242z. Epub 2007 Oct 31.
In an effort to identify the new resistance factors in rifampin resistant (RIF (r)) Mycobacterium tuberculosis ( M. tb), comparative proteome analysis and gene mutation assays were used to identify the differentially expressed proteins and correlated gene mutations among clinical RIF (r) isolates lacking rpoB mutations, RIF sensitive (RIF (s)) isolates, and the laboratory H37Rv strain. MALDI-TOF-MS revealed nine differentially expressed protein spots. PCR sequencing results showed four genes were mutated. The newly identified 191A/C mutation, in the gene Rv2629, was carried by 111 out of 112 clinical RIF (r) isolates. However, this mutation was absent in H37Rv and RIF (s) isolates. The RIF (s) species Mycobacterium smegmatis displayed RIF resistance only after being transformed with the mutated M. tb Rv2629, while it was not restored by the wild type gene. These results indicate that the 191A/C mutation of the Rv2629 gene may be associated with RIF resistance.
为了鉴定耐利福平(RIF(r))结核分枝杆菌(M. tb)中的新耐药因子,采用比较蛋白质组分析和基因突变检测方法,以鉴定缺乏rpoB突变的临床RIF(r)分离株、RIF敏感(RIF(s))分离株和实验室H37Rv菌株之间差异表达的蛋白质和相关基因突变。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)显示有9个差异表达的蛋白质斑点。PCR测序结果显示有4个基因发生了突变。新鉴定出的Rv2629基因中的191A/C突变,在112株临床RIF(r)分离株中有111株携带。然而,H37Rv和RIF(s)分离株中不存在这种突变。耻垢分枝杆菌RIF(s)菌株仅在用突变的结核分枝杆菌Rv2629转化后才表现出对利福平的耐药性,而野生型基因不能使其恢复。这些结果表明,Rv2629基因的191A/C突变可能与利福平耐药性有关。