牛lncRNA SNHG12过表达载体构建  

Construction of LncRNA SNHG12 Overexpression Vector

作　　者：郭虹 郝琴琴 李鹏飞[2] GUO Hong;HAO Qin-qin;LI Peng-fei(Animal Husbandry Development Center of Fanshi County,Fanshi,Shanxi 034300;College of Life Science,Shanxi Agricultural University,Taigu,Shanxi 030801)

机构地区：[1]繁峙县畜牧业发展中心,山西繁峙034300 [2]山西农业大学生命科学学院,山西太谷030801

出　　处：《中国牛业科学》2023年第2期6-10,共5页China Cattle Science

基　　金：山西省应用基础研究计划面上项目(20210302123380);山西农业大学校地合作项目(2021HX23,2022HX010);山西省现代农业产业技术体系建设专项。

摘　　要：可卡因苯丙胺调节转录肽(cocaine-and amphetamine-regulated transcript,CART)是一种内源性神经肽类物质,CART广泛存在于牛卵巢中,并且对牛卵泡闭锁产生调节作用。miRNA 491可以靶向结合牛CART基因3′-UTR区域,从而调控mRNA表达极显著下调。通过ENCORI数据库预测发现lncRNA SNHG12可能作为miRNA 491的分子海绵,从而影响牛CART基因的转录调控。本试验通过构建牛lncRNA SNHG12过表达重组载体,为进一步探究lncRNA SNHG12、miRNA 491和牛CART基因三者的互作关系奠定试验基础。结果显示pcDNA3.1-EGFP-SNHG12载体序列检测正确;转染293T细胞转染效果良好。荧光定量PCR结果显示在293T细胞中NC组的lncRNA SNHG12的表达水平极显著高于miRNA 491组,试验表明pcDNA3.1-EGFP-SNHG12载体构建成功,并且lncRNA SNHG12与miRNA491具有互作作用。Cocaine-amphetamine regulated transcript(CART)is an endogenous neuropeptide substance.CART is widely found in bovine ovaries and has a regulatory effect on bovine follicular atreasis.miRNA 491 can bind to the 3'-UTR region of bovine CART gene,thereby regulating mRNA expression significantly down-regulated.It has predicted that lncRNA SNHG12 may act as a molecular sponge of miRNA 491 based on ENCORI database,thereby affecting the transcriptional regulation of bovine CART genes.In this study,bovine lncRNA SNHG12 overexpression recombinant vector was constructed,which would lay a foundation for further exploration of the interaction between lncRNA SNHG12,miRNA 491 and bovine CART genes.The results showed that the sequence of pcDNA3.1-EGFP-SNHG12 vector was correct.The transfection effect of 293T cells was good.Fluorescence quantitative PCR results showed that the expression level of lncRNA SNHG12 in the NC group was significantly higher than that in the miRNA 491 group.The experiment showed that the pcDNA3.1-EGFP-SNHG12 vector was successfully constructed,and lncRNA SNHG12 interacted with miRNA491.

关 键 词：pcDNA3.1-EGFP-SNHG12 荧光定量技术 CART 细胞转染 

分 类 号：Q782[生物学—分子生物学]