Expression of SETD4 in Bac-to-Bac baculovirus expression system
Chinese Journal of Clinical Anatomy ›› 2015, Vol. 33 ›› Issue (4) : 426-429.
Expression of SETD4 in Bac-to-Bac baculovirus expression system
Objective To express SET domain-containing 4 (SETD4) protein through using baculovirus expression system and purify the expressed product to explore the functions of SETD4 protein and further understand the biological roles of SET family proteins. Methods The SETD4 gene was amplified by RT-PCR from mouse normal liver tissue. The gene was then inserted into pFastBac-HTB vector to form the recombinant donor plasmid which was further transformed into DH10Bac to construct the recombined bacmid. Next the bacmid was transfected to sf9 cells for package of the recombinant baculovirus particles. The recombinant SETD4 protein was expressed from the cells transduced by the recombinant baculovirus and was purified by NI-NTA resin. Purified protein was examined by coomassie brilliant blue staining and Western Blotting. Results The donor plasmid and recombined bacmid were successfully prepared and the recombinant baculovirus particles were produced from sf9 cells. The SETD4 protein was obtained and confirmed by brilliant blue staining and western blotting with a His-tag antibody and a specific SETD4 antibody. Conclusions The recombinant SETD4 protein was successfully expressed and prepared through baculovirus expression system.
SETD4 protein / Baculovirus expression system / Expression / Purification
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