Main content area

Efficient gene tagging Arabidopsis thaliana using a gene trap approach

Babiychuk, E., Fuangthong, M., Montagu, M. van., Inze, D., Kushnir, S.
Proceedings of the National Academy of Sciences of the United States of America 1997 v.94 no.23 pp. 12722-12727
Arabidopsis thaliana, genetic transformation, insertional mutagenesis, complementary DNA, structural genes, transgenic plants, Agrobacterium radiobacter, plasmid vectors, gene expression, gene targeting, nucleotide sequences, reporter genes, mutants, phosphotransferases (kinases)
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.