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Vol 45(2011) N 3 p. 466-471; D.A. Shirokov1*, V.V. Ryabichenko1, R.I. Akishina1, T.P. Ospel'nikova2, A.V. Glazunov1, G.G. Chestukhina1, V.P. Veiko1 Development of Hybrid Human Interferon Alfa-2 Strain-Producers and the Use of Enteropeptidase for Production of N-Terminal Methionine-Free Interferons 1Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 117545 Russia2Gamaleya Scientific Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, Moscow, 123098 Russia *mielnir@gmail.com Received - 2010-09-21; Accepted - 2010-10-14 We developed a technique for production of human interferon-α2a (IFN-α2a) and IFN-α2b lacking the N-terminal methionine. First of all, we constructed plasmids containing genes of hybrid IFN-α2 controlled by different promoters; the proximal part of the genes incorporated various sequences encoding the enteropeptidase cleavage site. As a result, four strains of Escherichia coli producing hybrid IFN-α2 were obtained. The protocol for IFN-α2 renaturation, cleavege of its N-terminal part, and chromatographic purification of the N-terminal methionine-free IFN-α2 was developed. interferon- 2a, interferon- 2b, enteropeptidase, enterokinase |
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