Expression and Purification of Human Testis Spermatocyte Apoptosis-related Gene,Tsarg2 Protein in E.coli

Objective To obtain recombinant TSARG2 protein by prokaryotic expression for its functioned study.Methods The encoding sequence for mature peptide of human TSARG2 was amplified with RT-PCR and inserted into pQE30 vector to establish the prokaryotic expressing system coupled with 6×His.The competent cells of host strain of M15 were transformed by the recombinant plasmid(pQE30/TSARG2).Expression of the target protein was induced with IPTG and assayed by SDS-PAGE after sequencing.The recombinant products were purified by Ni 2+ -NTA agarose column.Results:The cloned fragment of human TSARG2 was 100% consistent with that in Genbank(AY040204),which was deduced to express 305 Aa mature peptide of human TSARG2 correctly.The expressed fusion protein was 35 kD in SDS-PAGE as expected and 66% in the total quatiy of germ proteins.Conclusion:The homologous recombinant human TSARG2 was obtained after Ni-affinity chromatograph,which could be used for its functioned study and preparation of specific antibodies.