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目的本研究通过诱导结核分枝杆菌在低氧环境中进入持留状态,来比较分析不同时间不同低氧环境下结核分枝杆菌中的PhoP基因和PhoR基因的表达水平差异,探讨研究PhoPR双组分信号转导系统对结核分枝杆菌持留性的调控机制。方法首先对结核分枝杆菌国际标准无毒株(H37Ra)和结核分枝杆菌国际标准强毒株(H37Rv)在不同低氧环境下进行培养,提取每个样本菌株的总RNA,并进行完整性鉴定;运用SYBR Green I实时荧光定量PCR检测每个样本菌株中PhoP基因和PhoR基因的表达水平;比较分析不同时间不同低氧环境下的结核分枝杆菌菌株PhoP基因和PhoR基因的表达水平差异。结果在不同时间点对低氧环境的结核分枝杆菌PhoP基因和PhoR基因的表达水平进行检测,结果显示:与第10d相比,培养至第15d时H37Rv菌株和H37Ra菌株中的PhoP基因和PhoR基因的表达水平均显著上调,差异有统计学意义(P<0.05);并且培养至第25d时,H37Rv菌株中的PhoP基因和PhoR基因的表达水平比H37Ra菌株表达均上调2.34倍,差异有统计学意义(P<0.05)。结论在不同时间点相同毒力的结核分枝杆菌的PhoPR双组分信号转导系统中PhoP基因和PhoR基因在不同低氧环境下的表达存在差异,而且在相同时间点不同毒力结核分枝杆菌的PhoPR双组分信号转导系统中PhoP基因和PhoR基因在不同低氧环境下的表达也存在差异,表明PhoPR双组分信号转导系统对结核分枝杆菌的持留性具有调控作用。
Objective To study the difference of PhoP gene and PhoR gene expression between Mycobacterium tuberculosis and Mycobacterium tuberculosis under different hypoxia environment by inducing Mycobacterium tuberculosis to stay in hypoxia environment. Regulatory Mechanism of Signal Transduction System on Mycobacterium tuberculosis Persistency. Methods Mycobacterium tuberculosis isolates (H37Ra) and Mycobacterium tuberculosis (H37Rv) were cultured under different hypoxic conditions. The total RNA of each sample was extracted and the integrity . The expression levels of PhoP and PhoR genes in each sample were detected by SYBR Green I real-time fluorescence quantitative PCR. The differences of PhoP and PhoR gene expression between different strains of Mycobacterium tuberculosis under different hypoxia were compared. Results The levels of PhoP gene and PhoR gene in Mycobacterium tuberculosis were detected at different time points. The results showed that PhoP gene and PhoR gene in H37Rv and H37Ra strains cultured on the 15th day compared with the 10th day, (P <0.05). Moreover, the expression level of PhoP and PhoR genes in H37Rv strain was 2.34 times higher than that in H37Ra strain at the 25th day after culture, showing statistically significant difference Significance (P <0.05). Conclusion The PhoPR two-component signal transduction system of Mycobacterium tuberculosis with the same virulence at different time points showed different expression of PhoP gene and PhoR gene in different hypoxic environment, and different virulence tuberculosis branches at the same time point Bacillus PhoPR two-component signal transduction system PhoP gene and PhoR gene expression in different hypoxia environment there are differences, indicating that PhoPR two-component signal transduction system of Mycobacterium tuberculosis retention has a regulatory role.