一种改良的鉴定原核生物非编码RNA的方法

An improvement approach of identification of small RNAs in prokaryote

目的:构建一种鉴定原核生物非编码RNA(又称sRNA)的方法模型。方法:以鼠疫耶尔森菌为模型,基于非编码RNA的cDNA文库构建,改良RNA片段分离方法、获取全长RNA序列的RACE技术及rRNA去除方法,再应用RNA印迹法(Northern blot)对文库结果进行验证。结果:同目前常用的检测原核生物sRNA方法相比较,本改良方法更易于获得真实可信和全长的sRNA分子。结论:获得了一种较完整、片段覆盖范围较广的原核生物非编码RNA的鉴定方法。

Objective： To construct an improvement approach of identification small RNAs （sRNAs） in prokaryote. Methods： Taking Yersinia pestis as a model, constructed eDNA library by combining the RACE （rapid amplification of eDNA ends） technique with advanced RNA size selecting protocol and di- minishment of rRNAs, and finally verified the results using Northern blot. Results： Compared with cur- rently methods, the new constructed method was inclined to gain authentic sRNAs with full length and rich abundance. Conclusion： An improved method of eDNA library construction was prepared success- fully, by which more full-length prokaryotic sRNAs can be acquired.