DSpace Repository

Characterization of promoter tagged lines

Show simple item record

dc.contributor.author Remy, Serge
dc.contributor.author Thiry, Else
dc.contributor.author Windelinckx, Saskia
dc.contributor.author Rymen, Bart
dc.contributor.author Santos Ordoñez, Efren German
dc.contributor.author Coemans, Bert
dc.contributor.author Swennen, Rony
dc.contributor.author Sagi, Laszlo
dc.date.accessioned 2009-04-04
dc.date.available 2009-04-04
dc.date.issued 2004-07
dc.identifier.citation Remy S., Thiry E., Windelinckx S., Rymen B., Santos E., Coemans B., Swennen R. and Sági L., 2004. Characterization of promoter tagged lines. 1st International Congress on Musa. Harnessing research to improve livelihoods. Abstract guide. Penang, Malaysia, 6-9 July 2004. 29. Oral Abstract. en
dc.identifier.uri http://www.dspace.espol.edu.ec/handle/123456789/4824
dc.description.abstract We have developed a fast, sensitive and high-throughput method for the identification and isolation in bananas of novel promoters and genes. Following the Agrobacterium-mediated transformation of embryogenic cell suspension cultures with a promoterless luciferase (luc) gene, candidate promoters are characterized in planta during the early stages of in vitro development. Screening for luciferase (LUC) enzymatic activity in transgenic cell colonies has been performed without induction treatments. We have previously reported that transformation of a luciferase reporter trap linked to the left T-DNA border resulted in an activation frequency of 0.03-0.07%. All tagged candidates have now been propagated for plant regeneration and molecular analysis. Southern hybridization and sequence analysis of several independent lines demonstrated the integration of 2-6 T-DNA copies in the genome with the frequent presence of the vector backbone or complex rearrangements. Therefore, TAIL-PCR products from selected tagged lines were individually cloned and sequenced. One tagged line displayed constitutive expression higher than that of the CaMV 35S promoter, which persisted in the regenerated plants, both in vitro and in the greenhouse. Detailed analysis of the LUC expression in all plant tissues will be presented. One of the T-DNA insertions in this line occurred in a metallothionein-like protein gene, demonstrating the feasibility of the technique with bananas. An improved tagging construct, which contains the codon-optimized luc+ gene fused close to the right T-DNA border, has been prepared. Transformation with this construct resulted in a 40-fold increase in activation frequency (up to 2.5% without induction) compared to the original tagging construct. Data on LUC expression in hundreds of independent tagged lines will be presented. This T-DNA trapping technology is currently being applied to isolate inducible genes and promoters. en
dc.description.sponsorship Laboratory of Tropical Crop Improvement, Katholieke Universiteit Leuven. en
dc.language.iso eng en
dc.publisher 1st International Congress on Musa. Harnessing research to improve livelihoods. en
dc.rights openAccess
dc.title Characterization of promoter tagged lines en
dc.type Presentation en

Files in this item

This item appears in the following Collection(s)

Show simple item record

Search DSpace

Advanced Search


My Account