Construction of fibroblast activation protein expression vector and expression

Objective To construct an E.coli system expressing fibroblast activation protein (FAP) in orderto obtain the whole protein. Method Fibroblast activation protein cDNA was amplified by RT-PCR from humanfresh gastric carcinoma tissue, then cloned to vector pMD18-T. After the FAP sequence was confirmed, the FAPcDNA was isolated and inserted into expression vector pQE30. The recombinant plasmid pQE30-FAP wastransformed into JM109, then it was induced by Isopropyl-β-D-thiogalactopyranoside (IPTG) and FAP wasexpressed. Results The sequence of cloned FAP was identical with that published on GenBank. Expression vectorpQE30-FAP was constructed; after transformed into E.coli JM109, a whole protein was obtained, which was laterproven to be the induced protein of FAP. Conclusion We have successfully constructed the expression vector ofpQE30-FAP, from which whole protein FAP, proven by western blotting, is obtained. This research provides abasis to make AFP antibody.