Title: Improved T-DNA vector for tagging plant promoters via high-throughput luciferase screening
Authors: Remy, Serge ×
Thiry, Els
Coemans, Bert
Windelinckx, Saskia
Swennen, Rony
Sagi, Laszlo #
Issue Date: May-2005
Publisher: Eaton publishing co
Series Title: Biotechniques vol:38 issue:5 pages:763-770
Abstract: Transferred DNA (T-DNA) tagging is a powerful tool for tagging and in planta characterization of plant genes on a genome-wide scale. An improved promoter tagging vector is described here, which contains the codon-optimized luciferase (luc(+)) reporter gene 31 bp from the right border of the T-DNA. Compared to the wild-type luciferase gene, this construct provides significantly increased reporter gene expression and a 40 times higher tagging frequency. The utility of the construct is demonstrated in banana, a tropical monocot species, by screening embryogenic cell colonies and regenerated plants with an ultrasensitive charged-coupled device (CCD) camera. The improved vector resulted in a luciferase activation frequency of 2.5% in 19,000 cell colonies screened. Detailed molecular analysis of flanking DNA sequences in a tagged line revealed insertion of the luciferase tag in a novel gene with near-constitutive expression.
ISSN: 0736-6205
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Division of Crop Biotechnics
× corresponding author
# (joint) last author

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