Efficient gene tagging in Arabidopsis thaliana using a gene trap approach
AUTOR(ES)
Babiychuk, Elena
FONTE
The National Academy of Sciences of the USA
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=25099Documentos Relacionados
- Secretion Trap Tagging of Secreted and Membrane-Spanning Proteins Using Arabidopsis Gene Traps1
- Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana.
- Trichome Development in Arabidopsis thaliana. I. T-DNA Tagging of the GLABROUS1 Gene.
- Gene and Enhancer Trap Tagging of Vascular-Expressed Genes in Poplar Trees
- A genomics approach to the chaperone network of Arabidopsis thaliana