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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.