Construction of prokaryotic expression plasmid with rat Sirt 1 gene and fusion protein expression

Objective To construct a recombinant expression plasmid with rat Sirt1 cDNA and to obtain the fusion protein.Methods A fragment of rat Sirt1 cDNA containing BamHI/XhoI were amplified by RT-PCR.The fragment was cloned into pGEM-T easy vector and subcloned into pET41 vector.Subsequently,E.coli BL21 cells were transformed by the recombinant plasmid.The fusion protein,which was induced by IPTG was purified and was analyzed by SDS-PAGE.Results A fragment of 506 bp of rau Sirt1 cDNA was amplified by RT-PCR.The construction of the recombinant plasmid pET41-Sirt1 was confirmed by sequencing.The fusion protein of 48 kD was expressed and purified.Conclusions The recombinant plasmid has been successfully constructed.The fusion protein of rat Sirt1 can be expressed in E.coli BL21 cells with the expected molecular weight.