人巨细胞病毒miR-US33-1-5 p在感染的人胚肺成纤维细胞中靶mRNA的筛选与鉴定

摘要

目的 为了研究人巨细胞病毒 (human cytomegalovirus,HCMV) 编码的miR-US33-1-5p生物学功能,通过实验室方法筛选和鉴定出miRNA-US33-1-5p作用的靶mRNA.方法 利用一种简单有效的hybrid-PCR实验方法筛选HCMV miRNA-US33-1-5p的候选靶mRNA.并通过双荧光素酶检测系统鉴定 HCMV miRNA-US33-1-5p 对这些候选靶 mRNA 3′-UTR 的调控作用.结果 通过hybrid-PCR实验方法获得了HCMV miRNA-US33-1-5p在感染的人胚肺细胞中的12个候选靶mRNA.并证实HCMV miR-US33-1-5p使其中7个候选靶mRNA 3′-UTR的荧光素酶质粒表达荧光素酶的能力显著下降.结论 鉴定的miR-US33-1-5p靶mRNA编码的蛋白质功能涉及病毒复制、免疫系统调节、蛋白质合成、细胞周期调节、能量代谢等.HCMV miR-US33-1-5p 靶 mRNA 的鉴定为进一步研究HCMV miRNA-US33-1-5p的生物学功能提供实验资料.%Objective To screen and identify mRNAs targeted by human cytomegalovirus-encoded miR-US33-1-5p (HCMV miR-US33-1-5p) for understanding the biological functions of HCMV miR-US33-1-5p. Methods Potential target mRNAs of HCMV miR-US33-1-5p were screened out by using hybrid-PCR, a simple and effective method. Dual-Luciferase Reporter Assay System was used to identify the binding abili-ties of HCMV miR-US33-1-5p to 3′-UTRs of these potential target mRNAs. Results Twelve potential mRNAs targeted by HCMV miR-US33-1-5p were screened out in human embryo lung fibroblast cells infected with HCMV. Luciferase activities of 3′-UTRs of seven target mRNAs were significantly down-regulated. Conclusion Proteins encoded by these target mRNAs are involved in virus replication,immune system reg-ulation,protein synthesis,cell cycle regulation,energy metabolism and so on. Identification of mRNAs tar-geted by HCMV miR-US33-1-5p is helpful for further studies on biological functions of miR-US33-1-5p.