摘要
目的:通过免疫共沉淀和基因芯片技术,建立一种RNA 5'端帽子结构的鉴定方法。方法:利用m3G帽子结构的特异性抗体对线虫的总RNA进行免疫共沉淀,为线虫127个非编码RNA设计探针,制备了检测基因芯片,对其帽子结构进行鉴定,并对RNA的转录进行分析。结果:37个非编码RNA被分离鉴定为含有m3G帽子结构,与其它物种已发现m3G帽子的RNA比较,结果基本一致。结论:免疫共沉淀结合基因芯片技术对非编码RNA 5'端帽子结构分离鉴定的方法是可行有效的。此方法特异性强,灵敏度高,对大规模RNA帽子结构的鉴定、新RNA的发现和RNA功能的鉴定都具有一定的参考和应用价值。
Objective：To establish a protocol to determine the structure of RNA 5＇end cap by using co- immunoprecipitation （Co-IP） in conjunction with microarray technology. Methods： Total RNAs isolated from Caenorhabditis elegans were precipitated with the antibody against M3 G cap structure. For Microarray analysis, the probes were designed for 127 non-coding RNAs （neRNAs） of C. elegans. The cap structure was determined and RNA transcription was also analyzed. Results： 37 ncRNAs was isolated and identified as containing m3 G cap structure. This number is similar with other species, for which m3 G cap RNAs have been found. Conclusions ： It is feasible and effective to determine the structure of ncRNA 5＇ end cap by using Co-IP followed by Microarray analysis. This method is of great specificity and high sensitivity, which maybe applied to the large-scale RNA cap structure identification, the discovery of novel RNA and the identification of RNA functions.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2014年第2期65-70,共6页
China Biotechnology
基金
高等学校博士学科点专项科研基金(20111403110004)资助项目
关键词
帽子结构
鉴定
免疫共沉淀
基因芯片
RNA Cap structure Determination Immunopreeipitation Mieroarray
