Aim: To construct prokaryotic vector GX1-rmhTNF containing neovascular-targeting peptide GX1 and human TNFalpha and produce the GX1-rmhTNF protein.
Methods: We have previously obtained a specific phage peptide CGNSNPKSC (GX1) binding to vasculature of human gastric cancer. The GX1-rmhTNFalpha vector was constructed by merging sequence of neovascular-targeting peptide GX1 with N-terminal of new recombined human TNFalpha (rmhTNFalpha) using gene engineering methods. The expression of the fusion protein GX1-rmhTNFalpha in E. coli was induced by temperature. The expression of GX1-rmhTNFalpha was detected by SDS-PAGE and Western blot.
Results: A novel protein with expected molecular mass about 18,000 was found after SDS-PAGE and gel staining. The expressed product showed a good binding ability to anti-TNFalpha monoclonal antibody.
Conclusion: The prokaryotic vector GX1-rmhTNF was constructed and the expression of GX1-rmhTNF protein was successfully induced, which was helpful for further purification of GX1-rmhTNF protein.