Journal of Southern Medical University ›› 2013, Vol. 33 ›› Issue (12): 1757-.

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Construction of pQE/Dnajb13 recombinant plasmid and the protein expression

  

  • Online:2013-12-20 Published:2013-12-20

Abstract: Objective To construct pQE/Dnajb13 recombinant vector and induce the expression of the fusion protein. Methods
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 Ni2+ affinity chromatography. Western blotting was used to detect Dnajb13 expression. Results 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. Conclusion
We have successfully constructed pQE/Dnajb13 recombinant vector, which may facilitate further investigation of the role of
Dnajb13 in spermatogenesis.