Target selected insertional mutagenesis on chromosome IV of Arabidopsis using the En-I transposon system.
Reverse genetics using insertional mutagenesis is an efficient experimental strategy for assessing gene functions. The maize Enhancer-Inhibitor (En-I) transposable element system was used to develop an effective reverse genetics strategy in Arabidopsis based on transposons. To generate insertion mutations in a specific chromosomal region we developed a strategy for local transposition mutagenesis. A small population of 960 plants, containing independent I transpositions was used to study local mutagenesis on chromosome IV of Arabidopsis. A total of 15 genes, located on chromosome IV, were tested for I insertions and included genes identified by the European ESSA I sequencing programme. These genes were of particular interest since homologies to other genes and gene families were identified, but their exact functions were unknown. Somatic insertions were identified for all genes tested in a few specific plants. Analysis of these progeny plants over several generations revealed that the ability to generate somatic insertions in the target gene were heritable. These genotypes that show high levels of somatic insertions can be used to identify germinal insertions in the progeny.
Speulman
E
E
CPRO, Department of Molecular Biology, PO Box 16, 6700 AA, Wageningen, The Netherlands.
van Asperen
R
R
van der Laak
J
J
Stiekema
W J
WJ
Pereira
A
A
D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology; D002875:Chromosomes / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D008957:Models, Genetic; D016254:Mutagenesis, Insertional / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods
D017931:DNA Primers; D004251:DNA Transposable Elements; D018744:DNA, Plant