黑龙江八一农垦大学学报2023，Vol.35Issue(6)：95-101,7.DOI:10.3969/j.issn.1002-2090.2023.06.015
MAVS ΔTM基因真核表达载体的构建及其在293T细胞中的表达定位
Construction of Eukaryotic Expression Vector of MAVS ΔTM Gene and Expression Localization in 293T Cells
摘要
Abstract
Mitochondrial antiviral signaling proteins(mitochondria antiviral signaling protein,MAVS)played an important role in the antiviral immune response of vertebrates.In order to further study the role of amino acids in the transmembrane region of MAVS in virus infection of host cells,c-terminal transmembrane domain deletion mutants of MAVS gene(MAVS ΔTM)were constructed by genetic engineering method.Firstly,the MAVS ΔTM gene fragment was amplified by PCR technology and connected to the vector pcDNA3.0-Flag by seamless cloning technology.After enzyme digestion identification and sequencing results,it was confirmed that the MAVS ΔTM gene was successfully cloned into the vector pcDNA3.0-Flag,and the recombinant plasmid was named pcDNA3.0-Flag-MAVS ΔTM.The recombinant plasmid pcDNA3.0-Flag-MAVS ΔTM was transfected into human renal epithelial cells(HEK-293T),and the expression and localization of pcDNA3.0-Flag-MAVS ΔTM were detected by western blot and indirect immunofluorescence.The results showed that pcDNA3.0-Flag-MAVS ΔTM was successfully expressed in 293T cells and evenly distributed in cells,which would laid a foundation for further study on the function of amino acids in the transmembrane region of MAVS protein.关键词
真核表达载体/线粒体抗病毒信号蛋白/跨膜区氨基酸Key words
eukaryotic expression vector/MAVS/MAVS ΔTM分类
生物科学引用本文复制引用
杨超玥,李钰昌,郑亮,高鑫誉,曹宏伟,吴志军,张华,高振秋,肖莉杰..MAVS ΔTM基因真核表达载体的构建及其在293T细胞中的表达定位[J].黑龙江八一农垦大学学报,2023,35(6):95-101,7.基金项目
黑龙江省自然科学基金联合引导项目(LH2019C046) （LH2019C046）
学成人才科研启动经费(定向培养博士人员:XDB202007). （定向培养博士人员:XDB202007）
