【摘要】 目的:研究伤寒沙门菌核糖核酸酶G(RNase G)对非编码RNA(ncRNA)T3956胞内水平的影响。方法:利用自杀质粒介导的同源重组方法制备伤寒沙门菌RNase G基因(rng)缺陷变异株;利用重组质粒pBAD将rng导入rng缺陷变异株,构建rng缺陷回补株;通过实时定量PCR分别比较伤寒沙门菌野生株、rng缺陷变异株、回补株等在不同生长时期的ncRNA T3956水平。结果:成功制备伤寒沙门菌rng缺陷变异株、rng缺陷回补株和空质粒对照株;实时定量PCR结果表明,rng缺陷株中T3956的胞内水平较野生株有所升高,并且在对数中期和稳态期升高得更加明显,而回补株胞内T3956的水平又得到恢复。结论:伤寒沙门菌RNase G能够参与对胞内ncRNA T3956水平的调控,并且在细菌对数生长中期和稳态期作用更为明显。更多还原

【Abstract】 Objective: To investigate the influence of RNase G on the regulation of non-coding RNA(ncRNA) T3956 in Salmonella enterica serovar Typhi(S.Typhi).Methods: The rng deleted mutant of S.Typhi was prepared by the homologous recombination mediated by suicide plasmid;the rng complementary strain was generated by transferring the recombinant plasmid pBADrng into the rng deleted mutant;qRT-PCR was performed to analyze the level of non-coding RNA T3956 in the wild strain,the rng mutant strain,the complementary strain and the control strain at different growth phases.Results: The rng deleted mutant of S.Typhi,the rng complementary strain and the control strain were constructed successfully.The results of qRT-PCR revealed that the cellular level of T3956 was increased in the rng mutant comparing to the wild type strain,especially at mid-log phase and stationary phase,and the level of T3956 was restored in the rng complementary strain.Conclusion: RNase G was involved in the regulation of the ncRNA T3956 levels in S.Typhi,and played a more important role in the regulation at mid-log and stationary phase.更多还原