Cloning and characterization of a novel abscisic acid (aba)-induced hva22-like protein from triticum turgidum spp. dicoccoides in response to drought stress
Doğan, Esen (2010) Cloning and characterization of a novel abscisic acid (aba)-induced hva22-like protein from triticum turgidum spp. dicoccoides in response to drought stress. [Thesis]
Official URL: http://192.168.1.20/record=b1301609 (Table of Contents)
An HVA22-like protein was found to be differentially expressed in root tissue of wild emmer wheat (Triticum turgidum ssp. dicoccoides), under prolonged drought stress conditions. In this study we were able to clone and characterize the open reading frame of HVA22-like protein from root tissue of wild emmer wheat accession number TR39477, which was previously shown to be a drought-tolerant genotype. Sequence analysis indicated that HVA22-like protein product was a membrane protein and had four hypothetical transmembrane domains. Presence of the protein was shown by expressing it both in Escherichia coli and Saccharomyces cerevisiae and analyzing with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Localization of the protein in the cell was observed in S. cerevisiae utilizing a vector containing green fluorescent protein (GFP) gene. Results obtained using a confocal laser microscope indicated that the transformation of yeast cells with only the empty vector containing GFP gene yielded in a homogenous distribution of the GFP upon induction with galactose whereas the HVA22-like protein tagged with GFP was localized in the cell. Expression of GFP tagged HVA22-like protein was further confirmed with western blot analysis using mouse anti-GFP antibody. The work presented in this thesis was the first study to identify and characterize the HVA22-like protein and its protein product from Triticum turgidum ssp. dicoccoides.
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