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Updated: Jul 4, 2026
High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
High-level expression, purification, and characterization of the recombinant grass carp pituitary adenylate
Zhangzhi Wang1, Mu Yang, Xinyan Wang
1School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu.
Abstract:
Pituitary adenylate cyclase-activating polypeptide-38 (PACAP(38)) is a potent secretagog for growth hormone and gonadotropin in fish species. To obtain recombinant grass carp PACAP(38), its open reading frame was subcloned in pET32a(+) vector to express thioredoxin (Trx)-PACAP fusion protein in Escherichia coli BL21 (DE3). The resulting expression level of the thioredoxin-PACAP reached 36% of the total proteins, and more than 85% of fusion protein existed as soluble form. Using Ni(2+)-chelating affinity chromatography, 102 mg of Trx-PACAP(38) with a purity of 97% was obtained from 342 mg of crude proteins from a 1-liter culture of Escherichia coli. The purified Trx-PACAP specifically inhibited T98G human glioblastoma cell proliferation, but the fusion partner had no effect in this regard. Moreover, this inhibition was totally abolished by PACAP-specific antibody.
