Selection and prokaryotic expression of MUC1 mimic epitope / 第三军医大学学报

ABSTRACT

Objective To screen MUC1 antigen mimic epitope by phage display peptide library technology and to construct a recombinant plasmid expressing MUC1 antigen mimic epitope.Methods MUC1 mimic epitope was screened by Biopanning,DNA sequencing and amino acid sequence comparison.The gene was constructed into PET-31b(+) expression vector and expressed in Escherichia coli BL21(DE3) after transformation and induction by IPTG.The complete protein of the host bacteria was extracted for SDS-PAGE.The recombinant protein was purified by affinity chromatography on a Ni~(2+)-sepharose column and detected by Western blotting.Results The selected MUC1 mimic epitope could specifically combine with MUC1 monoclonal antibody.The recombinant expression vector PET-31b(+)-MUC1 was constructed successfully after the fusion protein was induced with IPTG.A specific protein band was shown on SDS-PAGE profile and detected by Western blotting.Conclusion An MUC1 mimic epitope was screened out.The epitope fusion protein was successfully expressed for the development of tumor vaccines targeting MUC1.