Construction of pQE/Dnajb13 recombinant plasmid and the protein expression / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To construct pQE/Dnajb13 recombinant vector and induce the expression of the fusion protein.</p><p><b>METHODS</b>The open reading frame (ORF) of Dnajb13 gene was amplified from mouse testis cDNA library by PCR. The products were digested by SacI and SalI and subcloned into pQE vector. After identification by DNA sequence analysis, the recombinant plasmids were transformed into competent E.coli M15 cells. The His/Dnajb13 fusion protein was expressed with IPTG induction and purified with Ni(2+) affinity chromatography. Western blotting was used to detect Dnajb13 expression.</p><p><b>RESULTS</b>The recombinant vector pQE/Dnajb13 was successfully constructed. His/Dnajb13 fusion protein was expressed abundantly at 4 h after IPTG induction, and Western blot analysis demonstrated the presence of Dnajb13 expression in E.coli M15.</p><p><b>CONCLUSIONS</b>We have successfully constructed pQE/Dnajb13 recombinant vector, which may facilitate further investigation of the role of Dnajb13 in spermatogenesis.</p>