Overlap PCR法克隆人LRRFIP1基因及其在人肝细胞中诱导干扰素的作用研究  

LRRFIP1 cDNA Clone Through Overlap PCR and Its Role in the Induction of Interferon in Human Hepatocytes

作　　者：刘媛[1] 王文博[2] 邹自英[1] 朱冰[1] 胡宗海[1] 熊杰[1] 

机构地区：[1]成都军区总医院检验科,中国四川成都610083 [2]成都军区疾病预防控制中心,中国四川成都610021

出　　处：《生命科学研究》2016年第6期492-496,共5页Life Science Research

基　　金：成都军区总医院院管课题基金(2013YG-B055);国家自然科学基金项目(81301445);全军医学科技青年培育项目(14QNP057)

摘　　要：富亮氨酸重复序列(Fli-1)相互作用蛋白-1(leucine-rich repeat(in flightless I)interacting protein-1,LRRFIP1)可与人flightless I(FLI)蛋白的leucine-rich repeat(LRR)结构域结合,具体作用并不明确,可能参与肌动蛋白的组装。近年来研究发现LRRFIP1在小鼠成纤维细胞和巨噬细胞中能够促进Ⅰ型干扰素的表达,同时LRRFIP1的过表达能够降低病毒感染水平,提示LRRFIP1可能参与病毒感染诱导的先天免疫应答。肝脏作为人体重要的消化器官,容易受到乙型肝炎病毒和丙型肝炎病毒等的感染,现通过重叠延伸PCR(overlap PCR)方法,从肝癌细胞株Huh7中成功克隆了人LRRFIP1基因,并在肝细胞内成功过表达,发现Huh7细胞内LRRFIP1的过表达可上调干扰素(interferon,IFN)-β的表达水平,这为后续深入研究LRRFIP1基因的生理功能提供了一定的参考数据。Leucine-rich repeat（in flightless I） interacting protein-1（LRRFIP1） was originally identified as a protein that interacts with the leucine-rich repeat（LRR） domain of human flightless-I.Its biological role is unknown,and might be important for actin organization.Recent studies have found that LRRFIP1 can promote the expression of type I interferon in mouse fibroblasts and macrophage cells.Over-expressed LR-RFIP1 reduced the levels of virus infection,suggesting that LRRFIP1 may be involved in viral infection-induced innate immune response.Liver,as an important human digestive organ,is vulnerable to hepatitis B virus and hepatitis C virus infection.Here,human LRRFIP1 gene was successfully cloned from hepatoma Huh7 cells via overlap PCR method.It was found that over-expression of LRRFIP1 in Huh7 cells up-regu-lated the expression of interferon（IFN）-β.The results may provide additional information for the physiological functions of human LRRFIP1.

关 键 词：富亮氨酸重复序列(Fli-1)相互作用蛋白-1(LRRFIP1) 重叠延伸PCR 基因克隆 肝细胞 I型干扰素 

分 类 号：Q781[生物学—分子生物学]