Proceedings of the XLVI Italian
Society of Agricultural Genetics - SIGA Annual Congress
Giardini
Naxos, Italy - 18/21 September, 2002
ISBN 88-900622-3-1
Poster
Abstract - 3.29
GENE-TRAPPING IN ARABIDOPSIS
LANDONI M., DE FRANCESCO A., GODI M.,
MORONI B., PIGOZZI S., PROCISSI A., SOLIGON S., TRIOLO D., GALBIATI M.
Dipartimento di Genetica e di Biologia
dei Microrganismi, Università degli Studi di Milano, Italia
Arabidopsis
thaliana, transposable element, gene-trapping, GUS pattern
The
completion of the A. thaliana genome sequence showed the presence
of 25,498 predicted genes, but only 9% of the genes have been experimentally
characterized, and about 70% of the genes have only a putative function.
Our
group is a member of a network working on the Exotic (Exon Trapping Insert
Consortium) project, founded through the Fifth Framework programme of the EU
for Plant Biotechnology.
The
goal of this projects is to initiate a large-scale programme aimed at
determining the expression patterns of approximately 5,000 genes of Arabidopsis by
the generation of a new population of 50,000 transposon insertions in the Arabidopsis
genome.
The
system used in this project is based on the maize Ac and Ds
transposable element. The Ds elements carry the b-glucuronidase
(GUS) gene as a reporter and the Neomycin phosphostransferase (NPTII) gene
(conferring resistance to kanamycin) as a selectable marker.
Random
insertion of the Ds element throughout the genome allow us to detect
chromosomal gene expression through the activation of the GUS gene.
Mutagenesis is
initiated by crossing plants homozygous for one of the Ds elements
to plants containing the Ac transposase gene. The double
resistant F2 seedlings, containing a transposed Ds
element, are allowed to self-fertilize and the resulting F3 plants are stained
for GUS activity and examined for mutant phenotypes.
We will present
data about the generation of this collection, the GUS expression patterns
obtained, and the results of a preliminary characterisation of some transposant
lines.