title: Efficient gene tagging in Arabidopsis thaliana using a gene trap approach.
abstract: Large quantities of DNA sequence information about plant genes are rapidly accumulating in public databases, but to progress from DNA sequence to biological function a mutant allele for each of the genes ideally should be available. Here we describe a gene trap construct that allowed us to disrupt transcribed genes with a high efficiency in Arabidopsis thaliana. In the T-DNA vector used, the expression of a bacterial reporter gene coding for neomycin phosphotransferase II (nptII) depends on the in vivo generation of a translation fusion upon the T-DNA integration into the Arabidopsis genome. Analysis of 20 selected transgenic lines showed that 12 lines are T-DNA insertion mutants. The disrupted genes analyzed encoded ribosomal proteins (three lines), aspartate tRNA synthase, DNA ligase, basic-domain leucine zipper DNA binding protein, ATP-binding cassette transporter, and five proteins of unknown function. Four tagged genes were new for Arabidopsis. The results presented here suggest that gene trapping, using nptII as a reporter gene, can be as high as 80% and opens novel perspectives for systematic gene tagging in A. thaliana.
authors.affiliation: Laboratorium voor Genetica, Departement Genetica, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Universiteit Gent, B-9000 Gent, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Babiychuk
authors.forename: M
authors.initials: M
authors.lastname: Fuangthong
authors.forename: M
authors.initials: M
authors.lastname: Van Montagu
authors.forename: D
authors.initials: D
authors.lastname: Inzé
authors.forename: S
authors.initials: S
authors.lastname: Kushnir
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D018744:DNA, Plant; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional
chemical_list: D018744:DNA, Plant