畜牧与兽医2018，Vol.50Issue(6)：68-73,6.
利用MS2识别序列标签纯化技术富集猪链球菌小RNA结合蛋白
A purification technique for adding MS2 recognition sequence tag to RNA to enrich Streptococcus suis small RNA binding proteins
摘要
Abstract
Bacterial RNA-binding proteins not only play a key role in regulation of small RNAs,but also participate in regulation of bacterial virulence.However,reports on identification and function of bacterial RNA-binding proteins are rare in gram-positive bacteria.In order to establish an ideal method for screening Streptococcus suis RNA-binding proteins,S.suis small RNAs rss04 and rss06 were selected.The MS2-MBP that binds to the MS2 sequence was purified;the plasmids pSET2-MS2-rss04 and pSET2-MS2-rss06,expressing the fusion RNAs MS2-rss04 and MS2-rss06 respectively,as well as the negative control plasmid pSET2-MS2-negative,were constructed and then electroporated into the S.suis small RNA deletion mutants Δrss04,Δrss06,and wild strain P1/7,respectively.Then,the MS2 recognition sequence tag purification technology was used,and RNA was extracted from eluent.Quantitative real-time PCR results showed that rss04 and rss06 were significantly enriched by 16 folds and 110 folds,respectively,expressing fusion RNAs in the samples,whereas rss04 and rss06 were not enriched in the negative control group.The above results indicated that the MS2 purification system was successfully established and could be used for screening S.suis RNA-binding proteins,providing a technical platform for in-depth study on the function of S.suis small RNA.关键词
猪链球菌/RNA结合蛋白/小RNA/MS2标签Key words
Streptococcus suis/RNA-binding protein/small sRNA/MS2 tag分类
农业科技引用本文复制引用
张寿明,唐欢宇,吴宗福..利用MS2识别序列标签纯化技术富集猪链球菌小RNA结合蛋白[J].畜牧与兽医,2018,50(6):68-73,6.基金项目
国家自然科学基金项目(31572544) （31572544）
