Construction, Expression And Purification Of Human Myoglobin Protein

Aim: To obtain recombinant human myohaemoglobin protein (H-Mb) by prokaryotic expression for clinical use in diagnosis.Methods: The encoding sequence for mature peptide of human H-Mb 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-Mb). Expression of the target protein was induced with IPTG and assayed by SDS-PAGE and immunoblot after sequencing. The recombinant products were purified by Ni2+-NTA agarose column.Results: The cloned fragment of human H-Mb was 98.5% consistent with that in genbank (NM203378), which was deduced to express 153 animo acids mature peptide of human H-Mb correctly. The expressed fusion-protein was 16.7kD in SDS-PAGE as expected and was recognized by a commercial McAb.Conclusion: The homologous recombinant human H-Mb is obtained after Ni2+-NTA affinity chromatograpH, which may benefit for preparation of specific antibodies and diagnosis for some related diseases.