TY - CHAP
T1 - In vitro translation to study HIV protease activity
AU - Matsuda, Zene
AU - Iga, Mutsunori
AU - Miyauchi, Kosuke
AU - Komano, Jun
AU - Morishita, Kazuhiro
AU - Okayama, Akihiko
AU - Tsubouchi, Hirohito
PY - 2007/4/4
Y1 - 2007/4/4
N2 - 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.
AB - 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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U2 - 10.1385/1-59745-388-9:135
DO - 10.1385/1-59745-388-9:135
M3 - Chapter
C2 - 17634600
AN - SCOPUS:34547653036
SN - 1597453889
SN - 9781597453882
T3 - Methods in Molecular Biology
SP - 135
EP - 149
BT - In Vitro Transcription and Translation Protocols
A2 - Grandi, Guido
ER -
