Efficient gene tagging Arabidopsis thaliana using a gene trap approach

• Published in: Proceedings of the National Academy of Sciences - PNAS 1997-11, Vol.94 (23), p.12722-12727
• Main Authors: Babiychuk, E, Fuangthong, M, Montagu, M. van, Inze, D, Kushnir, S
• Format: Article
• Language: English
• Subjects: ADN; AGROBACTERIUM TUMEFACIENS; Arabidopsis - genetics; ARABIDOPSIS THALIANA; Biological Sciences; Biology; Cellular Biology; COMPLEMENTARY DNA; Deoxyribonucleic acid; DNA; DNA, Plant; Exons; EXPRESION GENICA; EXPRESSION DES GENES; Flowers & plants; GENBANK/Z86093; GENBANK/Z86094; GENBANK/Z86095; GENE; GENE DISRUPTION; GENE EXPRESSION; Gene Transfer Techniques; GENES; Genes, Plant; GENETIC TRANSFORMATION; Genetic Vectors; GENETICA MOLECULAR; GENETIQUE MOLECULAIRE; Genomes; Genomics; Homologous recombination; INSERTIONAL MUTAGENESIS; KINASES; Life Sciences; MOLECULAR GENETICS; MOLECULAR SEQUENCE DATA; MUTACION; Mutagenesis, Insertional; MUTANT; MUTANTES; MUTANTS; MUTATION; NEOMYCIN PHOSPHOTRANSFERASE II; NPTII GENE; NUCLEOTIDE SEQUENCE; Phenotypes; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; PLASMID VECTORS; REPORTER GENES; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; STRUCTURAL GENES; T-DNA VECTORS; TRANSFERASAS; TRANSFERASE; TRANSFERASES; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; TRANSGENIC PLANTS
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.94.23.12722

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