结核分支杆菌H_(37)Rv与H_(37)Ra株mRNA基因差异显示研究

Study on mRNA differential display in Mycobacterium tuberculosis H_(37)Rv and H_(37)Ra

目的 采用mRNA差异显示技术研究H3 7Rv株与H3 7Ra株的基因差异 ,为获取结核分支杆菌毒力相关基因奠定基础。方法 提取结核分支杆菌H3 7Rv株和H3 7Ra株RNA ,经逆转录、PCR扩增、聚丙烯酰胺凝胶电泳和放射自显影进行mRNA的差异显示。回收差异条带、再扩增和序列分析 ,进行同源性查询。结果 经差异显示后 ,共回收与H3 7Ra株不同的H3 7Rv株mRNA差异条带 10条。经结果查询 ,发现其中 2条差异条带与结核分支杆菌H3 7Rv株DNA序列具有同源性 ,而与H3 7Ra无同源性。结论 mRNA差异显示方法具有高效、灵敏等优点 ,可在基因水平掌握同一种属或不同种属不同株间的差异 ,用于结核分支杆菌相关毒力基因的研究。研究获得的H3 7Rv株与H3

Objective To study genetic differences between H 37 Rv and H 37 Ra by mRNA differential display, paving the way for obtaining virulence related gene of Mycobacterium tuberculosis (MTB).Methods Total RNA was extracted from MTB H 37 Rv and H 37 Ra, mRNA differential display was conducted following reverse transcription, polymerase chain reaction, polyacrilamide gel electrophoresis and radiography. The differential bands were recovered, reamplified, sequenced, and then homologous query was done.Results 10 differential bands between H 37 Rv and H 37 Ra were recovered, 2 of which were homologous to H 37 Rv DNA after query from NCBI database of GenBank.Conclusions mRNA differential display is an effective, sensitive method, can display differences between different strains intra and extra species and be used in searching Mycobacterium tuberculosis virulence related gene. Obtained differential bands in this study may cover MTB virulence related gene.