The cloning of the mouse BALB/c metallothionein-1 cDNA in the plasmid expression vector pNZ2103 of Lactococcus

Objective To clone the mouse BALB/c metallothionein-1(MT-1)cDNA in the plasmid expression vector pNZ2103 of Lactococcus.Methods Two primers were designed with the digestion site of Pst 1 and of Hind Ⅲ.With the plasmid pET21a-MT used as the template,MT-1 cDNA was amplified by PCR.MT-1 cDNA was cloned into pNZ2103 with the restriction endonuclease Pst Ⅰ and Hind Ⅲ to form the recombinant plasmid pNZ2103-MT.pNZ2103-MT was transformed into the strain DH5α.The transformant containing the plasmid pNZ2103-MT was identified by PCR and digestion of Pst I and Hind Ⅲ.The metallothionein expression capacity of pNZ2103-MT in DH5α was identified on 12% SDS-PAGE.Results The transformant(DH5α) containing the plasmid pNZ2103-MT was obtained.Conclusion MT expression of pNZ2103-MT in DH5α was identified on 12% SDS-PAGE.It was difficult to decide MT expression level because the amount was too low.