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目的研究结核分枝杆菌假设蛋白Rv0341编码基因iniB的多态性,确认其保守性,分析依赖利福平结核分枝杆菌(依R菌)在利福平(R)培养管中Rv0341高表达的可能原因。方法用大肠埃希菌做为阴性对照,提取依R菌(14株)、耐R菌(23株)、R敏感的结核分枝杆菌(12株)、结核分枝杆菌标准株H37Rv及BCG(共51株)的细菌基因组DNA,利用PCR自基因组DNA中扩增iniB基因,通过ApaⅠ/EcoRⅤ、KpnⅠ/SmaⅠ双酶切,琼脂糖凝胶电泳检测其带型,对iniB基因进行限制性片段长度多态性分析并测序。结果实验组中iniB基因的酶切图谱完全相同,测序结果亦证实iniB基因无突变。结论 iniB基因在对R依赖、耐药和敏感等不同类型的结核分枝杆菌中是相对保守的;依R菌在Rv0341蛋白表达方面的差异可能是RNA转录差异或利福平诱导的结果。
Objective To study the polymorphism of iniB gene encoding Rv0341 in Mycobacterium tuberculosis and to confirm the conservation of Rv0341 gene in Rifampin (R) Possible Causes. Methods Escherichia coli was used as a negative control and 14 strains of R bacteria, 23 strains of R bacteria, 12 strains of R-sensitive Mycobacterium tuberculosis, H37Rv strains of Mycobacterium tuberculosis and BCG A total of 51 isolates were obtained. The iniB gene was amplified from genomic DNA by PCR, and the iniB gene was amplified by restriction endonuclease digestion with ApaⅠ / EcoRⅤ, KpnⅠ / SmaⅠand agarose gel electrophoresis. The restriction fragment length Polymorphism analysis and sequencing. Results The iniB gene in the experimental group had the same digestion map, and the sequencing result also confirmed that there was no mutation in the iniB gene. Conclusion The iniB gene is relatively conserved in different types of Mycobacterium tuberculosis such as R, drug resistance and susceptibility. According to the expression of Rv0341 in R gene, the difference in RNAi may be the result of RNA transcription difference or rifampicin induction.