Construction of Arabidopsis P4H-7 GUS reporter and fluorescent tagging cassettes for Arabidopsis transformation
2006
Bou Daher, F.
The synthesis of hydroxyproline in plants and animals requires the post-translational hydroxylation of peptidyl-proline catalyzed by prolyl-4 hydroxylase (P4H) enzymes. Hydroxyproline is also found in mammalian hypoxia-inducible factor (HIF), a transcription factor and participates in the regulation of protein stability and consequently in hypoxic adaptation. In plants, there is no information on P4H involvement in hypoxic and anoxic response. To this direction, we develop molecular tools to study their spatial and temporal expression under oxygen deficiency and during plant growth and development. The Arabidopsis P4H-7 promoter was fused to GUS cDNA and the construct was transformed in Arabidopsis plants using vacuum infiltration. Several independent transformants were identified and their characterization is under way. In addition, the Fluorescent Tagging of Full Length Protein (FTFLP) technique was used to label the P4H7 gene with the gene of the cyan variant of the yellow fluorescent protein (CFP). The construct introduction into a gateway vector and the transformation of Arabidopsis plants is in progress. These molecular tools will be used to elucidate the physiological role of P4Hs in plant growth and development.
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