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.
国家科技期刊平台
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维普
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