Construction of fibroblast activation protein expression vector and expression

Objective: To construct an E.coli system expressing fibroblast activation protein (FAP) in order to obtain the whole protein. Method: Fibroblast activation protein cDNA was amplified by RT-PCR from human fresh gastric carcinoma tissue, then cloned to vector pMD18-T. After the FAP sequence was confirmed, the FAP cDNA was isolated and inserted into expression vector pQE30. The recombinant plasmid pQE30-FAP was transformed into JM109, then it was induced by Isopropyl-β-D-thiogalactopyranoside (IPTG) and FAP was expressed. Results: The sequence of cloned FAP was identical with that published on GenBank. Expression vector pQE30-FAP was constructed; after transformed into E.coli JM109, a whole protein was obtained, which was later proven to be the induced protein of FAP. Conclusion: We have successfully constructed the expression vector of pQE30-FAP, from which whole protein FAP, proven by western blotting, is obtained. This research provides a basis to make AFP antibody.