Soluble expression and preliminary study on a functional peptide of FnBP protein mediating Staphylococcus aureus invasion
AIM: to express the gene fragment of interesting peptide that comprised of C-terminal D1 and N-terminal D2 (FnBP-D1c/D2n)from the genome of 04018 in E. coli in GST-fusion form by use of DNA recombinant technique and genetic engineering. METHODS: The specific primers were synthesized to extract the fragment of interesting peptide from the genome of the staphylococcus aureus, then the gene fragment of interesting peptide was correctly cloned into expression vector pGEX-KG. The recombinant plasmid pGEX-KG was introduced into E. coli DH5α that was introduced to express the protein in GST-fusion form. RESULTS: Reconstructed gene could be expressed effectively in E. coli in soluble form. The molecular weight of expressed product of recombinantion plasmid pGEX-KG analyzed by SDS-PAGE was the same as anticipated. And the soluble expresssion product accounted for 20% of the total bacterial protein. ELISA assay showed that the expression product could bind to fibronectin. CONCLUSION: The expression vector has been constructed and reconstructed gene was expressed successfully and effectively in E. coli, which may provide a useful basis on developing new therapeutic and vaccine strategies for S. aureus invasion of cells.
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