P.93K-T QDPR Lentivirus Gene Expression Vector Construction

Objectives Construct dihydrogen biological disc poison reductase (quinoiddihydropteridine reductase, QDPR) and p.93K-T expression lentivirus vectors, forQDPR p.93K-T function research provides a powerful tool.Methods Respectively to extract contains QDPR and its mutant genes’ mRNA inrats,and reverse transcription for cDNA, using molecular cloning technology the twogene will be insert lentivirus vectors pLVX-IRES-ZsGreen1. PLVX-IRES-ZsGreen1, QDPR and p.93K-T recombinant plasmid respectively with psPAX2andpMD2G were co transfected into293T cells respectively, for empty, QDPR and p.93K-TQDPR express lentivirus vectors. The harvested lentivirus vectors were used to infectNRK-52E cells respectively, the infected cells were observed by fluorescencemicroscopy, the expression of QDPR protein in each group was used for detection ofwestern blot.Results Empty, QDPR, p.93K-T QDPR Lentivirus Expression under fluorescencemicroscopy showed green fluorescence, the infection rate of close to100%. Western blotresults showed that QDPR, QDPR protein content in p.93K-T was significantly higherthan that in normal group and empty vector group.Conclusion Successfully to construct the lentiviral vector QDPR, p.93K-T QDPR,provide a powerful tool for studying the function of p.93K-TQDPR.