Expression of human lysozyme in Pichia pastoris mediated by intein-ELP

To study the expression of human lysozyme in Pichia pastoris mediated by intein and Elastin-like polypeptide(ELP),the hLYZ-intein-ELP gene was cloned into the yeast secretory expression vector pPIC9Kto construct the recombinant plasmid pPIC-MIELYZ.Then the recombinant plasmid was transformed into P.pastoris GS115after digested by restriction enzyme Pme I,positive clones were screened with G418resistance for expression induced by 0.5% methanol.SDS-PAGE and Western blot confirmed the correct molecular weight of recombinant protein,antibacterial assay showed that recombinant protein had obvious antimicrobial activity against Micrococcus.These results show that the fusion gene has expressed successfully in P.pastoris,and lay the foundation for Subsequent purification work and construction of new Pichia pastoris expression vector.