Construction of plant gene expression vectors carrying ath-Mir393a gene and its target gene AFB3
Bakar, Mine (2010) Construction of plant gene expression vectors carrying ath-Mir393a gene and its target gene AFB3. [Thesis]
Official URL: http://192.168.1.20/record=b1302879 (Table of Contents)
MicroRNAs (miRNA) are a class of non-coding small RNAs that regulate gene expression. Plant miRNAs are involved in a broad range of biological processes including growth, developmental patterning and abiotic stress responses. In our previous miRNA profiling results in response to glyphosate, ath-Mir393a was found to be up-regulated at the 5 % glyphosate application whereas down-regulated at the 20 % foliar glyphosate application when compared to control samples of Festuca arundinacea. Ath-Mir393a is a conserved miRNA species that targets F-box proteins TIR1 and AFB1/2/3 transcripts. F-box motif mediates protein-protein interactions and mostly found in proteins as components of SCF ubiquitin-ligase complexes. Hence they have very important roles in the maintenance of protein stabilities inside the cell. TIR1 and AFBs have shown that they are associated with auxin signaling pathways as auxin receptors. In order to improve the management of weed invasion on crop lands, the idea of developing genetically engineered cultivars which are glyphosate-resistant has become a necessity. Therefore in this study two plant gene expression vectors carrying Arabidopsis thaliana ath-Mir393a gene and AFB3 gene were constructed by using Gateway technology. Ath-Mir393a and AFB3 genes were first captured in 'entry vectors' and then recombined into pEARLEY Gate100 Gateway compatible 'destination vectors'. These vectors are later used for Agrobacterium-mediated transformation of Arabidopsis thaliana. These vectors constructed in this study will be a good starting point for exploring and elucidating the miRNAs associated with regulation of plant responses to glyphosate.
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