Abstract
The overexpression of a gene in a heterologous system is often the prelude to or the prerequisite of the elucidation and characterization of a given protein, in particular where the protein is difficult to obtain in sufficient quantity from natural sources. Prokaryotic expression systems, in particular Escherichia coli have been exploited successfully for a number of viral proteins. A large number of E. coli expression systems are currently commercially available that offer the investigator a large number of choices with regard to promoter choice, site of expression, fusions, and so forth. Since the variety and number of expression systems available are extensive, it is not within the scope of this chapter to discuss them fully, and the final choice of expression system used, is often arrived at empirically and often reflects the investigator’s “favored system.”
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References
Hijikata, M, Mizushima, H, Akagi, T., Mori, S., Kakiuchi, N., Kato, N, et al. (1993) Two distinct protease activities required for the processing of a putative non-structural precursor protein of Hepatitis C Virus. J. Virol 67, 4665–4675
Grakoui, A, McCourt, D W, Wychowski, C, Feinstone, S M, and Rice, C M (1993) A second Hepatitis C virus-encoded proteinase. Proc Natl Acad Sci USA 90, 10,583–10,587.
D’Souza, E. A., O’Sullivan, E., Amphlett, E M, Rowlands, D. J, Sangar, D V., and Clarke, B. E. (1994) Analysis of NS3-mediated processing of Hepatitis C virus non-structural region in vitro. J Gen Virol 75, 3469–3476.
Grakoui, A, McCourt, D W, Wychowski, C., Feinstone, S. M., and Rice, C. M. (1993) Characterisation of Hepatitis C virus-encoded serine proteinase: Determination of proteinase-dependent polyprotein cleavage sites J Virol 67, 2832–2843
Bartenschlager, R., Ahlborn-Laake, L., Mous, J, and Jacobsen, H. (1993) Nonstructural protein 3 of the Hepatitis C virus encodes a serine-type proteinase required for cleavage at the NS3/4 and NS4/5 Junction J Virol 67, 3835–3844.
Bartenschlager, R., Ahlborn-Laake, L., Yasargil, K, Mous, J, and Jacobsen, H (1995) Substrate determinants for cleavage in cis and trans by the Hepatitis C virus NS3 proteinase J Virol 69, 198–205
Lin, C, Pragai, B M, Grakoui, A, Xu, J, and Rice, C M (1994) Hepatitis C virus NS3 serine proteinase, trans-cleavage requirements and processing kinetics. J. Virol 68, 8147–8157
Han, D S, Hahm, B., Rho, H, and Jang, S K (1995) Identification of the protease domain in NS3 of Hepatitis C virus. J. Gen Virol 76, 985–993.
Bartenschlager, R, Lohmann, V, Wilkinson, T., and Koch, J O (1995) Complex formation between the NS3 serine-type proteinase of the Hepatitis C virus and NS4A and its importance for polyprotein maturation J Virol 69, 7519–7528
Lin, C. and Rice, C M. (1995) The Hepatitis C virus NS3 serine proteinase and NS4A cofactor: Establishment of a cell-free trans-processing assay. Proc Natl Acad Sci. USA 92, 7622–7626
Failla, C, Tomei, L, and De Francesco, R (1994) Both NS3 and NS4A are required for proteolytic processing of Hepatitis C virus nonstructural proteins. J Virol 68, 3753–3760.
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© 1998 Humana Press Inc., Totowa, NJ
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D’Souza, E., Gray, N., Ellis, M., Clarke, B.E. (1998). Expression and Characterization of HCV NS3 Protease. In: Lau, J.YN. (eds) Hepatitis C Protocols. Methods in Molecular Medicine™, vol 19. Humana Press. https://doi.org/10.1385/0-89603-521-2:343
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DOI: https://doi.org/10.1385/0-89603-521-2:343
Publisher Name: Humana Press
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