Cloning, Expression and Purification of Human Heart Type Fatty Acid Binding Protein in E.coli

Objective To obtain recombinant human heart type fatty acid binding protein(H-FABP) by prokaryotic expression for clinical use in diagnosis. Methods The encoding sequence for mature peptide of human H-FABP was amplified with RT-PCR and inserted into pQE30 vector to establish the prokaryotic expressing system coupled with 6His. The competent cells of host strain of M15 were transformed by the recombinant plasmid(pQE-H-FABP). Expression of the target protein was induced with IPTG and assayed by SDS-PAGE and immunoblot after sequencing. The recombinant products were purified by Ni~+-NTA agarose column. Results The cloned fragment of human H-FABP was 99.9% consistent with that in genbank(NM004012), which was deduced to express 132 Aa mature peptide of human H-FABP correctly. The Expressed fusion-protein was 15 kD in SDS-PAGE as expected, was recognized by a commercial McAb(Clone 6B6) and was 27% in the total quatiy of germ proteins. Conclusion The homologous recombinant human H-FABP is obtained after Ni~+-affinity chromatograph, which may benefit for preparation of specific antibodies and diagnosis for some related diseases.