CLONING, EXPRESSION AND PURIFICATION OF ABO3 GENE INVOLVED IN DROUGHT STRESS TOLERANCE FROM Arabidopsis thaliana in Escherichia coli

Abstract

       Drought is one of the most severe environmental stress factors that affects the growth and development of plants. The effects of drought are foreseen to increase with climate change and growing water scarcity. In many parts of the world, including Vietnam, plants may frequently encounter drought stress. Plants have evolved specific mechanisms to respond to drought stresses. Studying these protective mechanisms will contribute to our knowledge of tolerance and resistance to stress. Recently, some studies indicated that the ABO3 gene coding for WRKY63 is a key gene in stress signalling pathways related to drought tolerance, of which the functions and interactions are currently unknown. Therefore, investigating its functions and interactions are considered effective strategies to control a plant’s resistance against drought. In this study, we show the results of expression and purification of the ABO3 protein coding for WRKY63 in E. coli, a solid step towards studying about the protein’s interactions and functions. The ABO3 gene was amplified from cDNA of Arabidopsis thaliana by PCR, cloned into T-Blunt vectors, and sequenced. This gene then was subcloned into the expression vector pQE-30 and expressed in E. coli strain M15. Finally, the ABO3 recombinant protein was initially purified by affinity chromatography for further studies.