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Expression of the Japanese encephalitis virus NS3 and NS2b proteins as glutathione S-transferase fusions

Shivashankar, Y and Satchidanandam, V (1995) Expression of the Japanese encephalitis virus NS3 and NS2b proteins as glutathione S-transferase fusions. In: Indian Journal of Biochemistry & Biophysics, 32 (6). pp. 356-360.

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Flaviviruses generate their structural and nonstructural proteins by proteolytic processing of a single large polyprotein precursor. These proteolytic events are brought about both by host cell signalase and a virally encoded protease. The virally encoded proteolytic activity has been shown to reside within the nonstructural protein 3 (NS3) and requires the product of the nonstructural 2b (NS2b) gene. In order to obtain sufficient quantities of pure NS2b and NS3 proteins for kinetic analysis, we have expressed both these proteins in recombinant systems as fusions to glutathione S-transferase (GST). The fusion constructs were driven by the strong bacteriophage T7 promoter. Transfection of these constructs into the African green monkey kidney cell line CV-1 previously infected with a recombinant vaccinia virus expressing the T7 RNA polymerase resulted in synthesis of the fusion proteins. Both the fusion proteins could be purified to homogeneity in a single step using a glutathione agarose affinity matrix.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to National Institute of Science Communication and Information Resources.
Department/Centre: Division of Interdisciplinary Research > Centre for Nano Science and Engineering
Depositing User: Ms V Mangala
Date Deposited: 19 May 2011 04:20
Last Modified: 22 Oct 2018 10:34
URI: http://eprints.iisc.ac.in/id/eprint/37732

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