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Luciferase
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+ I& L$ E! L, E' @Luciferases are enzymes that emit light. A diverse group of organisms use luciferase-mediated bioluminescence to startle predators or to attract prey or mates. The luciferase from the North American firefly releases green light during the oxidation of its chemical substrate, luciferin. Other organisms, including plants, that express the luciferase (LUC) gene will also glow faintly green when supplied with luciferin. 2 r3 w# c. w; ?; v: J
2 z: q2 \1 _7 V5 ?! w" k1 [: RThe glow is widely used as an assay for LUC expression, which acts as a "reporter" for the activity of any regulatory elements that control its expression. Luciferase is particularly useful as a reporter in living cells and organisms. LUC gene fusions provide a "window" on to the mechanisms that regulate the activity of specific genes, in specific, living cells. Our interest is in the mechanism of rhythmic regulation in higher plants: a rhythmically-expressed luciferase reporter is one of our principal tools. Images of bioluminescence from our CAB UC Arabidopsis plants are shown at the top of this page (the cotyledons of two seedlings) and on the front page (an older, rosette plant). / D$ I* x- L! j9 U' i5 |
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The LUC+ gene from Promega Corporation is an order of magnitude brighter in Arabidopsis and has no disadvantages in other species, so we now use this routinely. The protein is cytoplasmic, whereas the native LUC is peroxisomal.
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/ q, ^8 [( u; q( x; {More information about: [Circadian Rhythms] [Video Imaging] [Arabidopsis]
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Luciferase Links 2 B+ j. q: s S6 o9 N& C9 G7 ~
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Bioluminescence in nature: pictures of luminescent organisms from Harbor Branch Oceanographic Institution, and from the Hastings lab, Harvard.
5 p4 b/ n# _# ]. r% Y: N5 A For a more recent review of techniques in plants, see "The Use of the Luciferase Reporter System for in Planta Gene Expression Studies" Plant Mol. Biol. Reporter vol. 18(2), 143 (2000).* S5 s, Y8 b3 j5 s7 `7 e$ o& u
Wessel Van Leeuwen, Marc J.M. Hagendoorn, Tom Ruttink, Remco Van Poecke, Linus( L5 \' W* H g
H.W. Van Der Plas and Alexander R. Van Der Krol. Link to 2.5 Mb PDF
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+ t+ d/ R R% r/ ~/ `Firefly Luciferase as a Reporter for Plant Gene Expression Studies. (Promega Notes, 1993)& G! E) e$ p3 ^, s% \4 @; e& H
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Video Imaging of Regulated Firefly Luciferase Activity in Transgenic Plants (Promega Notes, 1994)
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6 \& N) z+ R' w, g3 ~Luminometry for In Vivo and In Vitro Reporting of Firefly Luciferase (Promega Notes, 1993)
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# x0 ]9 Y1 l) X, Q+ q( cMichael White's Lab, Liverpool. (firefly luciferase as a reporter gene in mammalian cells)
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Jeremy Tavar駳 Lab, Bristol. (firefly luciferase as a reporter gene in mammalian cells)
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$ P) X% G* D [% R8 XAndrew Millar Home Page |
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