Cloning and expression of human lysozyme in Pichia Pastrois and identification of the activity

Objective Lysozyme cDNA of human(hLYZ) was amplified by PCR and cloned into a vector pMD18-T.Methods The positive plasmid contained human lysozyme cDNA was determinded by restriction enzyme analysis and PCR.The cDNA encoding hLYZ was subcloned into pPIC9k and transferred into Pichia Pastrois GS115 by electroporation.It proved that the positive recombinant plasmid pPIC9k-hLYZ contained cDNA encoding human lysozyme by restriction enzyme analysis and sequence determination.Results In the positive transformants,the human lysozyme was expressed as a fusion protein in Pichia GS115,which was verified by SDS-PAGE.Bacteriostatic activity was verified by Micrococcus lysodeikticus lysis test.Conclusion The evident bacteriostatic action was verified.