Summary
HIV-1 is an etiological agent of AIDS. One of the targets of the current anti-HIV-1 combination chemotherapy, called highly active antiretroviral therapy (HAART), is HIV-1 protease (PR), which is responsible for the processing of viral structural proteins and, therefore, essential for virus replication. Here, we describe an in vitro transcription/translation-based method of phenotyping HIV-1 PR. In this system, both substrate and PR for the assay can be prepared by in vitro transcription/translation. Protease activity is estimated by the cleavage of a substrate, as measured by enzyme-linked immunosorbent assay (ELISA). This assay is safe, rapid, and requires no special facility to be carried out. Our rapid phenotyping method of HIV-1 PR may help evaluate drug resistance, useful when choosing an appropriate therapeutic regiment, and could potentially facilitate the discovery of new drugs effective against HIV-1 PR.
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© 2007 Humana Press Inc., Totowa, NJ
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Matsuda, Z. et al. (2007). In Vitro Translation to Study HIV Protease Activity. In: Grandi, G. (eds) In Vitro Transcription and Translation Protocols. Methods in Molecular Biology™, vol 375. Humana Press. https://doi.org/10.1007/978-1-59745-388-2_7
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DOI: https://doi.org/10.1007/978-1-59745-388-2_7
Publisher Name: Humana Press
Print ISBN: 978-1-58829-558-3
Online ISBN: 978-1-59745-388-2
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