کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2089384 1545790 2006 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Firefly luciferase terminally degraded by mild heat exposure: Implications for reporter assays
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Firefly luciferase terminally degraded by mild heat exposure: Implications for reporter assays
چکیده انگلیسی

Luciferase reporter constructs are an accurate method of assessing gene promoter activity and vectors constitutively expressing luciferase are useful in quantifying transfection efficiency. Common methodologies for examining the induction of the heat shock (stress) response require exposure of cells transfected with luciferase-expressing vectors to a mild heat stress. Here we re-examine the under-recognised phenomenon that luciferase is exquisitely sensitive to small temperature changes. In cells subjected to mild heat exposure following transfection with both luciferase and β-galactosidase reporter vectors, a marked reduction in luciferase activity was observed compared with β-galactosidase activity. On exposing recombinant firefly luciferase to small increases in temperature in vitro, a time and temperature dependent decrease in luciferase activity was demonstrated. Loss of luciferase activity following mild heat exposure will result in misinterpretation of reporter activity. This vastly underappreciated effect is worthy of further emphasis and luciferase reporter vectors should be used with caution in protocols that involve exposure to temperatures outside the physiological range.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 310, Issues 1–2, 20 March 2006, Pages 182–185
نویسندگان
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