Construction of the prokaryotic expression vector and expression of protein phosphatase LY1 gene

Objective To construct a prokaryotic expression vector for LY1 protein phosphatase gene,and to express the gene in E.coli BL21(DE3).Methods LY1 gene fragment was amplified by PCR and cloned into the pET28a(+)vector.The recombinant vector was identified by restriction endonuclease digestion analysis and DNA sequencing,and then transformed into E.coli BL21(DE3)by IPTG induction to express the target protein.The recombinant protein was detected by SDS-PAGE and Westernblot.Results A 1 179 bp of LY1 gene fragment was obtained.After recombinant vector pET28a(+)-LY1 was transformed into E.coli BL21(DE3)and induced by IPTG induction,the recombinant protein with MW about 42 kD was obtained.SDS-PAGE analysis showed that the expressed product was mainly in inclusion bodies.Conclusions Recombinant expression vector pET28a(+)-LY1 is constructed successfully.The expressed LY1 protein will be helpful to our further research.