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Construction of targeting vectors to disruptPAT1 gene inArabidopsisTrp1-100

Research paper by Seonhee Lim, Youngsoon Kim, Euiseung Lee, Hyeonsook Cheong

Indexed on: 01 Sep '97Published on: 01 Sep '97Published in: Journal of Plant Biology



Abstract

To develop the gene targeting system by homologous recombination inArabidopsis thaliana, we constructed two targeting vectors and showed the reliability of the scheme which is based on genetic complementation of phosphoribosylanthranilate transferase (PAT1) gene. ThePAT1 gene, which is essential for tryptophan biosynthesis, was selected as a target gene because the loss of function leads to fluorescence phenotype due to the accumulation of anthranilic acid derivatives. pHS113 containsPAT1 gene surrounding 5′ and 3′ flanking portions, but the most coding region of thePAT1 gene is replaced by the neomycin phosphotransferase gene (NPTII). pHS117 consists of 1.1 kb internal fragment of genomicPAT1 gene following withNPTII gene. In this targeting strategy,Arabidopsis PAT1 gene can be disrupted by single-step of transformation experiment.