EXPRESSION VECTOR CONTAINING OUTER MEMBRANE PROTEIN C (OMPC) GENE DERIVED FROM E. COLI AS A CELL SURFACE ANCHORING MOTIF

摘要

PURPOSE: An expression vector containing OmpC gene as a cell surface anchoring motif, organisms transformed therewith and protein expression method on the cell surface with the transformed organism are provided. The vector is used such as to remove heavy metals by expression of poly-histidine peptide truncated with OmpC gene. CONSTITUTION: Chromosome DNA of E. coli MC4100 is used as a template to amplify OmpC gene from E. coli. 1.1 kb of PCR product is isolated from agarose gel and cloned into pTrc99A having strong trc promoter. The resulting plasmid pTrcC has two Pst1I at 7th loop and C-terminal. PstI site of C-terminal is removed to make cloning easily and the constructed plasmid is named as pTCdP. The resulting plasmid is transformed into E. coli MC4100 and E. coli MC 4100/pTCdP (KCYC0576BP) is obtained. Poly-histidine linker peptide (poly-His) consisted 6 histidine is amplified by PCR and cloned into recombinant expression vector pTCdP. pTCHP1 is obtained. Other expression vectors having 2,3 and 6 unit of poly-His are constructed. SDS-PAGE analysis shows that poly-His protein consists 30-40% of outer membrane protein.