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。
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.
出处
《江苏大学学报（医学版）》
CAS
2012年第2期114-119,共6页
Journal of Jiangsu University：Medicine Edition
基金
江苏省高校自然科学基金资助项目(10KJB310001)
