中国免疫学杂志2013，Vol.29Issue(6)：634-638,5.DOI:10.3969/j.issn.1000-484X.2013.06.017
IFITM2的克隆表达及其抗病毒作用初探
Cloning and expression of IFITM2 gene and preliminary study on its antiviral activity
摘要
Abstract
Objective:To construct the recombinant plasmid containing ifitm2 gene and detect its expression in HEK293T cells,and to investigate its antiviral activity.Methods:The primers of ifitm2 gene were designed and synthesized according to its gene sequence published on Genbank.Then ifitm2 gene was obtained by RT-PCR and subcloned to pMD18-T simple vector.After sequencing,the PCR product was inserted into the eukaryotic expression vector pVAX1,resulting into pV-IFITM2,which was transfected into HEK293 cells.The expression of IFITM2 was detected by RT-PCR,Western blot and laser scanning confocal microscope (LCSM).Simuhaneously,VSV-G protein-pseudotype virus containing EGFP was used to infect HEK293T cells over-expressed IFITM2 protein.The antiviral activity of IFITM2 was analyzed by Western blot.Results:The ifitm2 gene was obtained and inserted into pVAX1,and the recombinant plasmid containing ifitm2 gene was constructed successfully.The results of Western blot and LSCM showed that IF-ITM2 was expressed in HEK293 cells and distributed ubiquitously.The result of expression of EGFP contained VSV-G pseudotype virus showed that IFITM2 inhibited the infection of VSV-G pseudotype virus in a certain extent.Conclusion:The recombinant eukaryotic expression plasmid containing ifitm2 gene was successfully constructed and the effective expression of IFITM2 protein in HEK293 cells was accomplished,which could restrict the VSV-G pseudotype virus infection.This study provided the experimental and theoretical basis for the research of the antiviral effect and molecule mechanism of IFITM proteins.关键词
IFITM2/克隆表达/抗病毒作用Key words
IFITM2/Clone and expression/Antiviral activity分类
生物科学引用本文复制引用
李沂,李昌,杜寿文,王茂鹏,朱羿龙,刘存霞,秦艳青,金宁一..IFITM2的克隆表达及其抗病毒作用初探[J].中国免疫学杂志,2013,29(6):634-638,5.基金项目
本文为973计划前期研究专项课题(No.2011CB512110)和中国博士后科学基金面上项目(No.20110491803、No.2012T50859) （No.2011CB512110）
