Comparison of Vibrio and firefly luciferases as reporter gene systems for use in bacteria and plants

Mudge, Stephen R., Lewis-Henderson, Wendy R. and Birch, Robert G. (1996) Comparison of Vibrio and firefly luciferases as reporter gene systems for use in bacteria and plants. Australian Journal of Plant Physiology, 23 1: 75-83. doi:10.1071/PP9960075


Author Mudge, Stephen R.
Lewis-Henderson, Wendy R.
Birch, Robert G.
Title Comparison of Vibrio and firefly luciferases as reporter gene systems for use in bacteria and plants
Formatted title
Comparison of Vibrio and firefly luciferases as reporter gene systems for use in bacteria and plants
Journal name Australian Journal of Plant Physiology   Check publisher's open access policy
ISSN 1445-4408
1445-4416
0310-7841
Publication date 1996
Sub-type Article (original research)
DOI 10.1071/PP9960075
Volume 23
Issue 1
Start page 75
End page 83
Total pages 9
Place of publication Collingwood, VIC, Australia
Publisher CSIRO Publishing
Language eng
Formatted abstract
Luciferase genes from Vibrio harveyi (luxAB) and firefly (luc) were introduced into E. coli, Agrobacteriurn, Arabidopsis and tobacco. Transformed bacteria and plants were quantitatively assayed for luciferase activity using a range of in vitro and in vivo assay conditions. Both lux and luc proved efficient reporter genes in bacteria, although it is important to be aware that the sensitive assays may detect expression due to readthrough from distant promoters. LUX activity was undetectable by liquid nitrogen-cooled CCD camera assays on intact tissues of plants which showed strong luxAB expression by in vitro assays. The decanal substrate for the lux assay was toxic to many plant tissues, and caused chemiluminescence in untransformed Arabidopsis leaves. These are serious limitations to application of the lux system for sensitive, non-toxic assays of reporter gene expression in plants. In contrast, LUC activity was readily detectable in intact tissues of all plants with luc expression detectable by luminometer assays on cell extracts. Image intensities of luc-expressing leaves were commonly two to four orders of magnitude above controls under the CCD camera. Provided adequate penetration of the substrate luciferin is obtained, luc is suitable for applications requiring sensitive, non-toxic assays of reporter gene expression in plants.
Keyword LUC
LUX
Arabidopsis,
Tobacco
E. coli
Agrobacterium
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Agriculture and Food Sciences
 
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Created: Tue, 22 Mar 2011, 14:08:11 EST by Dr Stephen Mudge on behalf of Botany, Department of