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Production of functional rat liver PSP protein in Escherichia coli
T Oka1, Y Nishimoto, T Sasagawa
1Department of Veterinary Physiology, Faculty of Agriculture, Kagoshima University, Japan. oka@vet.agri.kagoshima-u.ac.jp
Abstract:
An efficient Escherichia coli expression system for the production of a perchloric acid-soluble protein (PSP) has been constructed. Complementary DNA encoding PSP was inserted into an inducible bacterial expression vector pGEX-4T-1. After the plasmid introduced into E. coli was expressed by isopropyl 1-thio-beta-D-galaetopyranoside (IPTG), the recombinant product was purified by glutathione-Sepharose 4B affinity chromatography. The purified product showed the expected NH2-terminal sequence, but the translation inhibitory activity of this product was 10 times lower compared with that of authentic PSP isolated from rat liver.