Construction of eukaryotic expression vector of human antimicrobial peptide LL-37 / 临床检验杂志

ABSTRACT

Objective To construct eukaryotic expression vector of human antimicrobial peptide LL-37 pPIC9-LL-37, and transform the plasmid pPIC9-LL-37 into P.pastoris GS115 to obtain the recombinant P.pastoris strains.Methods The full-length of antimicsobial peptide LL-37 gene was artificially synthesized by overlap extension method and was fused to pPIC9 and then the fused plasmid was transformed into E.coli DH5?.After analysis by PCR and sequencing,the plasmid pPIC9-LL-37 was transformed into P.pastoris.The colonies exhibiting the phenotype of His+Mut+ or His+Mut-were screened by means of MM and MD plates and the insertion was confirmed by PCR.Results The results of PCR and sequencing confirmed that the LL-37 gene was correctly inserted into pPIC9. The colonies of 10 His+Mut+ and 9 His+Mut-were obtained by means of MM and MD plates screening and were confirmed by PCR.Conclsion The recombinant P.pastoris strains containing LL-37 were successfully obtained.