کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3406741 1593487 2012 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A high-throughput assay for HIV-1 integrase 3′-processing activity using time-resolved fluorescence
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی ویروس شناسی
پیش نمایش صفحه اول مقاله
A high-throughput assay for HIV-1 integrase 3′-processing activity using time-resolved fluorescence
چکیده انگلیسی

HIV-1 integrase (HIV-1 IN), a well-validated antiviral drug target, catalyzes multistep reactions to incorporate viral DNA into the genome of the host cell; these include both a 3′-processing (3′P) reaction and a strand transfer reaction. These enzymatic activities can be measured in vitro with short DNA oligonucleotides that mimic a single viral LTR DNA end and purified IN. A highly sensitive and reproducible time-resolved fluorescence (TRF)-based assay for HIV-1 IN 3′P activity is now reported. This assay was optimized with respect to time and concentrations of metal ions, substrate and enzyme. The assay has now been used successfully to measure HIV-1 IN 3′P activity and has been shown to detect the anti-IN activity of several known 3′P inhibition compounds accurately. This assay, which is amenable to high-throughput screening, will be useful for identification of additional HIV-1 IN 3′P inhibitors.


► A time-resolved fluorescence-based assay for HIV-1 IN 3′ processing.
► Optimized for time and concentrations of metal ions, substrate and enzyme.
► Used to study IN 3′P kinetics and inhibition by both described and novel 3′P inhibition compounds.
► Highly sensitive and reproducible.
► Amenable for high-throughput screening.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 184, Issues 1–2, September 2012, Pages 34–40
نویسندگان
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