ANALYSIS OF Arabidopsis thaliana GAMMA CARBONIC ANHIDRASE 2 PROMOTER USING REPORTER GENES

MARTIN, M. V.; VILLARREAL, F.; EDUARDO JULIAN ZABALETA

Rosario

Congreso; 42th Reunion anual de SAIB; 2006

Arabidopsis thaliana ã-Carbonic Anhydrase 2 (AtãCA2) is a nuclear encoded member of a small protein family present in mitochondria. The submitochondrial localization of these proteins has been established as structural subunits of complex I (NADH:ubiquinone oxidoreductase) forming a plant-specific extra domain. Previous studies by in situ hybridization, and RT-PCR showed that gammaCA2 is expressed mainly in flowers, specifically in the tapetum of anthers. In this work, three AtãCA2 promoter deletions were fused to the reporter GUS gene. AtãCA2 promoter 2000 bp-, 1500 bp-, 500 bp- and 300 bp-GUS constructs were used to transform Arabidopsis by floral dip methods. Whole transgenic seedlings, adult leaves, roots and flowers were analysed for in situ GUS expression. Results showed that transcriptional activation of gammaCA2 promoter is affected in the different tissues and deletions analyzed, suggesting the presence of several tissue-specific regulatory elements. ã-Carbonic Anhydrase 2 (AtãCA2) is a nuclear encoded member of a small protein family present in mitochondria. The submitochondrial localization of these proteins has been established as structural subunits of complex I (NADH:ubiquinone oxidoreductase) forming a plant-specific extra domain. Previous studies by in situ hybridization, and RT-PCR showed that gammaCA2 is expressed mainly in flowers, specifically in the tapetum of anthers. In this work, three AtãCA2 promoter deletions were fused to the reporter GUS gene. AtãCA2 promoter 2000 bp-, 1500 bp-, 500 bp- and 300 bp-GUS constructs were used to transform Arabidopsis by floral dip methods. Whole transgenic seedlings, adult leaves, roots and flowers were analysed for in situ GUS expression. Results showed that transcriptional activation of gammaCA2 promoter is affected in the different tissues and deletions analyzed, suggesting the presence of several tissue-specific regulatory elements. ãCA2 promoter deletions were fused to the reporter GUS gene. AtãCA2 promoter 2000 bp-, 1500 bp-, 500 bp- and 300 bp-GUS constructs were used to transform Arabidopsis by floral dip methods. Whole transgenic seedlings, adult leaves, roots and flowers were analysed for in situ GUS expression. Results showed that transcriptional activation of gammaCA2 promoter is affected in the different tissues and deletions analyzed, suggesting the presence of several tissue-specific regulatory elements. ãCA2 promoter 2000 bp-, 1500 bp-, 500 bp- and 300 bp-GUS constructs were used to transform Arabidopsis by floral dip methods. Whole transgenic seedlings, adult leaves, roots and flowers were analysed for in situ GUS expression. Results showed that transcriptional activation of gammaCA2 promoter is affected in the different tissues and deletions analyzed, suggesting the presence of several tissue-specific regulatory elements.