Fox Hunting System: an application to auxin research.

Abstract

About 10,000 non-redundant Arabidopsis full length cDNA (fl-cDNA) clones (RAFL cDNA) were mixed to make a normalized fl-cDNA over-expression T-DNA library. Agrobacterium was transformed with the normalized T-DNA library to give the corresponding Agrobacterium library. A. thaliana Col-0 WT plants were dipped with the Agrobacteria library. About 15,000 transformed plants were grown in green house to isolate plants that showed obvious phenotypes such as morphological abnormality, greening and pigmentation. Among them four lines showed phenotype similar to plant over-expressing tryptophan 2-monooxigenase gene (auxin synthesis gene from bacteria). PCR was performed to amplify four different fl-cDNAs from these lines. These cDNAs were reintroduced into wild type plant to confirm recapitulation of the original phenotype. We designated this activation tagging-like system as Fox Hunting System (full-length cDNA over-expressor gene hunting system) to develop it as a novel technology for the functional genomics of agronomically important genomic materials.