伤寒沙门菌长链非编码RNA AS-RpoH分子鉴定

Identification of non-coding RNA AS-RpoH in Salmonella enterica serovar Typhi

目的:探讨伤寒沙门菌非编码RNA AS-RpoH的分子特性,为进一步研究其调控功能和机制奠定基础。方法:在AS-RpoH表达高峰区设计特异性探针,运用RNA印迹检测AS-RpoH的表达;通过PCR寻找AS-RpoH可能的转录起始位点和终止位点;在yhhk编码区设计特异性探针,运用RNA印迹分析yhhk的转录产物。结果:RNA印迹结果显示,AS-RpoH在沙门菌中确实存在,长约3 000 nt;PCR结果表明,AS-RpoH的转录终止位点位于rpo H基因起始密码子上游191~238 nt之间;而其转录起始位点可能位于yhh K上游;RNA印迹分析结果表明yhh K基因的转录产物与AS-RpoH的长度一致。结论:长链非编码RNA AS-RpoH在伤寒沙门菌中存在表达,可能为yhh K的3'非翻译区。

Objective： To clarify the molecular characteristics of a non-coding RNA,AS-RpoH in Salmonella enterica serovar Typhi（ S. Typhi）. Methods： Northern blotting analysis was used to investigate the expression of AS-RpoH in S. Typhi; RT-PCR was used to identify the possible transcription initiation and termination sites of AS-RpoH; Northern blotting analysis with specific probe was used to analyze the transcripts of yhhK. Results： The AS-RpoH expression was confirmed by Northern blotting with a length of approximate 3 000 nt; The transcription termination site of AS-RpoH was located in the region between 191-238 nucleotides of rpo H start codon upstream; The possible initiation site of AS-RpoH transcription was located on the upstream of yhhK; Northern blotting analysis revealed that the length of yhhK transcript was the same as the length of AS-RpoH. Conclusion： AS-RpoH is one of long non-coding RNAs expressed in S. Typhi,which may be a possible 3＇-untranslation region of the yhhK.