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Production of functional rat liver PSP protein in Escherichia coli |
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| Authors: | T. Oka Y. Nishimoto T. Sasagawa H. Kanouchi Y. Kawasaki Y. Natori |
| Affiliation: | (1) Department of Veterinary Physiology, Faculty of Agriculture, Kagoshima University, Kagoshima 890-0065 (Japan), JP;(2) Department of Nutrition, School of Medicine, University of Tokushima, Tokushima 770-8503 (Japan), JP;(3) Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197 (Japan), Fax +81 99 285 8714, e-mail: oka@vet.agri.kagoshima-u.ac.jp, 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-β-D-galactopyranoside (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. Received 8 October 1998; received after revision 6 November 1998; accepted 6 November 1998 |
| Keywords: | . Perchloric acid-soluble protein recombinant PSP Escherichia coli translation. |
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