GABI-DUPLO Double Mutants
Donated by
- Bernd Weisshaar CeBiTec (Center for Biotechnology), Faculty of Biology, Bielefeld University
- Dario Leister Plant Molecular Biology Group, Botanik, Department Biologie I, Fakultät für Biologie , Ludwig-Maximilians-Universität München (LMU)
- Cordelia Bolle Plant Molecular Biology Group, Botanik, Department Biologie I, Fakultät für Biologie , Ludwig-Maximilians-Universität München (LMU)
- Dario Leister Plant Molecular Biology Group, Botanik, Department Biologie I, Fakultät für Biologie , Ludwig-Maximilians-Universität München (LMU)
Click here to view all 367 of these lines.
Description
Generation of double mutants using T-DNA mutagenised A. thaliana plants (accession Columbia) as parental lines.
The GABI-DUPLO project was initiated to generate a collection of A. thaliana double mutants. A significant proportion of the genome of Arabidopsis thaliana consists of paralogous regions. In total, A. thaliana contains more than 24,000 similar gene pairs. Because those genes often have overlapping or redundant functions, their functional characterisation by forward or reverse genetics requires the inactivation of both gene copies. We have generated such double mutants (DUPLO lines) as a resource that will be instrumental for the functional characterisation of these genes. SALK and GABI-Kat insertion lines which share the Columbia-0 background have been the starting material for the double mutants.
Gene pairs were selected that share high similarity in sequence and show no homology to a potentially existing third related gene (~2,500 pairs). Gene pairs with close genetic linkage (<= 7.5Mbp) were not considered for the generation of double mutants.Parental lines, with in most cases insertions in the coding region, were crossed, and the F1 generation was selfed. The F2 generation was segregated and the genotype was determined by PCR. Homozygous double mutants were isolated and in the next generation the genotype of homozygous double mutants was rechecked and plants were phenotypically evaluated in the greenhouse. If no DUPLO line could be identified in the F2 generation, the offspring of a F2 line, which was homozygous for one insertion and heterozygous for the other, was allowed to self and the segregating F3 generation was propagated on sucrose-containing media plates to allow heterotrophic growth, and genotyped by PCR. If the segregation of the F3 seeds also did not yield any homozygous double mutant, these pairs were declared homozygous absent and are likely lethal as a double mutant and will therefore be donated as homozygous/heterozygous seeds.
A detailed description of the lines is made available from the GABI-DUPLO site (www.gabi-kat.de/duplo). DUPLO lines will be distributed with their respective parental lines.
Funded by the German Plant Genomics Program of BMBF
Ordering Information
Each set contains the double mutant and the parents. Please order the complete sets as opposed to the individual stocks.
PLEASE NOTE: Parental lines can still be a segregating population. Double mutants should be rechecked before doing experiments (primer sequences are provided on the GABI-DUPLO web page).
A detailed description of the lines is made available from the GABI-DUPLO site www.gabi-kat.de/duplo. An example that shows which data are available from the DUPLOdb is the description of the #double mutant 195#. DUPLO lines will be distributed with their respective parental lines.
The URL for #double mutant 195# is: http://www.gabi-kat.de/db/duplopair.php?pair_id=195
Clicking on the NASC code link (number beginning with N) from the link above will take you to the detailed page of the double mutant at NASC where you can then order the set.
As soon as the GABI-DUPLO project have the NASC-IDs for the DMs (IDs for the set, the DM and the parents) in their LIMS, additional data will show up on the "pair details" page at their site.
Number of Double Mutants available at NASC (367)
Batches 4,5,6,7 - 178 Double Mutants plus their parental lines made available - 18 April 2013
Batch 3 - 118 Double Mutants plus their parental lines made available - 4 April 2013
Batch 2 - 66 double mutant lines plus their parental lines made available - 22 March 2013.
Batch 1 - 5 double mutant lines plus their parental lines made available - 5 November 2012.