中国免疫学杂志2012，Vol.28Issue(5)：440-443,4.DOI:10.3969/j.issn.1000-484X.2012.05.014
HIV-1包膜蛋白ENV慢病毒载体的构建及表达
Lentivirus construction and expression in outer membrane protein ENV of HIV-1
摘要
Abstract
Objective:To construct lentiviral vector including HIV-1 envelope protein ENV and observe the env gene expression in the infected human embryonic kidney cells HEK293T. Methods:A complete HIV-1 env gene was obtained by point mutations. The env gene was subcloned to the EcoR I and Xho I sites of pLVX-IRES-ZsGreen 1, the recombinant plasmid pLV-env was constructed and transfected into HEK293T by the lipofectamine transfection method. The target gene expression was detected by RT-PCR and Western blot. Simultaneously, the env gene expression was located with the technology of laser scanning confocal microscope. Re-SultS;HIV-l env gene was obtained; the recombinant lentiviral plasmid pLV-env was constructed; the exogenous genes was expressed by detection of RT-PCR and Western blot. Meanwhile, the env gene could be secreted onto the cell membrane after expression. Conclusioil:The recombinant lentiviral plasmids including HIV-1 envelope protein env gene was constructed successfully, and its expression was verified. It laid foundation for the next lentiviral packaging and the construction of cell and animal models.关键词
ENV/HIV-1/慢病毒载体/真核表达Key words
ENV/HIV-1/Lentiviral vector/Eukaryotic expression分类
生物科学引用本文复制引用
孙丹丹,金宁一,李昌,李太元,朱娜,杜寿文,任大勇,刘存霞,秦艳青,李沂..HIV-1包膜蛋白ENV慢病毒载体的构建及表达[J].中国免疫学杂志,2012,28(5):440-443,4.基金项目
本文为国家自然科学基金(81001342),973计划(2011CB512110) （81001342）
吉林省自然科学基金(201015116),吉林省高新技术产业发展项目(2010)和长春市科技特派员行动计划(09KT04) （201015116）
