Construction and identification of mitochondrial antiviral signaling protein in eukaryotic expression vector

Abstract

Objective To construct the eukaryotic expression vector of mitochondrial antiviral signaling protein. Methods The target DNA fragments of mitochondrial antiviral signaling protein gene was obtained from PCR amplification, then the cDNA fragment was ligated into pMD™18-T Vector. After double-enzyme digestion, the MAVS gene was transferred into the eukaryotic expression vector pcDNA6/myc-HisA. The constructs transformed into E. coil JM109 and positive clones were picked by pcDNA6/myc-HisA-MAVS PCR amplification. This construct was confirmed by PCR and DNA sequencing. Results The eukaryotic expression vector of mitochondrial antiviral signaling protein was constructed correctly. The recombinant vector was transfected into OL cells, and the expression of the recombinant protein was detected by western blotting. Conclusion The successfully constructed pcDNA6/myc-HisA MAVS plasmid is useful to study the gene’s function and effects in cells.This study is for further study of the mechanism of gene function. Key words: Mitochondrial antiviral signaling protein; Eukaryotic expression vectors; Sequence analysis