pcDNA3.1-HisB-MEG3体外表达载体的构建及分析验证

pcDNA3.1-HisB-MEG3 Construction and Analysis of Expression Vector in Vitro

为了探究非编码RNA MEG3是否具有编码潜能,以及能否编码功能性微肽蛋白,本研究利用分子克隆技术克隆了牛的MEG3基因的预测ORF区核酸序列,并将其连入体外表达体系中,构建了pcDNA3.1-HisB-MEG3融合蛋白载体,获得MEG3的体外表达产物并验证其表达情况。结果表明:成功构建的pcDNA3.1-hisB-MEG3融合蛋白载体,无论是从RNA水平还是蛋白质水平,该融合蛋白均能够在外源的细胞中正常表达。综上可知,非编码RNA的MEG3是具有编码能力的,并且可以编码微肽,这一结果证实了MEG3的编码潜能,为后续探索MEG3生物学功能的研究夯实基础。

In order to explore whether non-coding RNA MEG3 has coding potential and whether it can encode functional micropeptide proteins. In this study, the nucleic acid sequence of predicted ORF region of bovine MEG3 gene was cloned by molecular cloning technology and connected into the in vitro expression system to construct pcDNA3.1-HisB-MEG3 fusion protein vector, obtained the in vitro expression product of MEG3 and verify its expression. The results showed that the successfully constructed pcDNA3.1-HisBMEG3 fusion protein vector could be normally expressed in foreign cells, both at RNA level and protein level. In conclusion, MEG3, a non-coding RNA, has the coding ability and can encode micropeptides, and which confirms the coding potential of MEG3 and lays a solid foundation for subsequent experimental studies exploring the biological functions of MEG3.