The Cloning And Expression Analysis Of ABI5Gene Promter From Arabidopsis Thaliana

Plant promter sequence contain many cis acting elements, it takes part in the regulation of the downsteam gene in the transcription level, thus, plant can take some adjustments to the external environment, to adapt the changes of the environment. Analysis of the promoter of one plant gene, could offer some experimental datas for the activation of its downstream gene expression. Abscisic acid (ABA) is an important hormone of the plants, it not only take part in many physiological process of plant, but also can regulate the metabolic processes of plant cells, to adjust the plants adapt to the environment. ABA-insensitice5(ABI5) is a basic leucine zipper protein which can response the ABA signal in seeds and vegetation organs. It can activate a series of gene that regulated by ABA, have important effections in the pathway of ABA signal, the expression of relevant genes, and embryonic development in which ways the ABI5regulate plants in its growth and development, and adjustment the changes of environment.Because of the Arabidopsis genome has been sequenced, we get the Arabidopsis ABI5promoter sequence from the NCBI gene bank data base directly. We analysised this sequence by bioinformatics method, the result shows that this sequence contain a lot of sequences that may associate with root and flower location and a quantity of cis acting elements, such as ABA-response elements, drought responsive elements, and light responsive elements, these possible sequences may act as some correspongding control effects of the exprence of ABI5gene. And there is a782bp intron before the transcription initiation site, it may effect the transcription activity of the promoter. In order to fully understand the specificity of exprence in time and space which the promoter sequence regulated, we designed a pair of primers, with these primers, we can get a sequence that include the whole promoter sequence and the untranslated regions(include the intron). And through the convention PCR, we get a1820bp sequence that end at the transcription initiation site (ATG), and test this sequence with the DNA sequencing. And then, we link this sequence with a double digested pCAMBIA1301plasmid to be an ABI5Promter::GUS recombinant plasmid. With the flral dipping method by agrobacterium tumefaciens, we integrated this recombinant gene into arabidopsis thaliana genome DNA, the screening and identification is in progress.