Cloning of mouse signal transducers and activators of transcription-4/6 and construction of eukaryotic expression plasimid / 重庆医科大学学报

ABSTRACT

Objective:To clone mouse signal transducers and activators of transcription(STAT) 4 and STAT6 gene and construct a eukaryotic expression plasimid.Methods:The cDNA of mouse STAT4 and STAT6 gene amplified with RT-PCR from normal mouse spleen tissue were cloned into pMD19-T vector by T/A ligation.After double digested,STAT4/STAT6 cDNA fragment was subcloned into pIRES2-EGFP vector to construct a eukaryotic expression plasimid.Restriction endonucleases analysis and sequencing were used to comform the recombinant plasimid.Results:The full length STAT4/STAT6 cDNA was correctly inserted into eukaryotic expression plasimid,and its sequencing was consistent with reported sequence derived from Genebank.Conclusion:The successful construction of eukaryotic expression plasimid provides a basis for futher studies on the infection of STAT4/STAT6 interaction to the downstream cytokines.