In vitro translation to study HIV protease activity

Zene Matsuda, Mutsunori Iga, Kosuke Miyauchi, Jun Komano, Kazuhiro Morishita, Akihiko Okayama, Hirohito Tsubouchi

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)


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.

Original languageEnglish
Title of host publicationIn Vitro Transcription and Translation Protocols
Subtitle of host publicationSecond Edition
EditorsGuido Grandi
Number of pages15
Publication statusPublished - Apr 4 2007

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics


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