Cloning of Human Thioredoxin Gene and Construction and Identification of Its Recombinant Retrovirus Vector

Objective To clone human thioredoxi(hTRX)gene and consturct recombinant retrovirus vector carrying the target gene.Methods The cDNA gene of human thioredoxin encoding protein was amplified by RT-PCR from PHA-stimulated human peripheral blood mononuclear cells(PBMCS).Then the amplification product was subcloned into retrovirus vector pSIV-1 identified by PCR,double endonuclease digestion and DNA sequencing.Results By comparing with the hTRX cDNA sequence reported in GenBank(BC003377),we found that the highly conserved enzymatic active site(Trp-Cys-Gly-Pro-Cys)of hTRX gene cloned was identical with that had been reported.However,there were four differences at bp132,136,170,264 and the corresponding amino acids of bp136,170 were changed(Phe→Leu,Ile→Thr).The pSIV-1-hTRX recombinant retrovirus vector has been obtained successfully.Conclusion Human thioredoxin gene cloning and construction of its recombinant retrovirus vector pSIV-1-hTRX paves the way for future study on biological activities and application of hTRX.