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Nghiên cứu tách dòng, biểu hiện và tinh sạch protein ABO3 từ cây Arabidopsis thaliana trong vi khuẩn Escherichia coli

Bài viết Nghiên cứu tách dòng, biểu hiện và tinh sạch protein ABO3 từ cây Arabidopsis thaliana trong vi khuẩn Escherichia coli trình bày Những ảnh hưởng của hạn hán được dự đoán là sẽgia tăng cùng với sự thay đổi khí hậu và làm trầm trọng hơn vấn đề khan hiếm nước sạch. Ở nhiều nơi trên thế giới bao gồm cả Việt Nam, thực vật thường xuyên phải đối mặt với vấn đề hạn hán,. . | Vietnam J. Agri. Sci. 2016, Vol. 14, No. 7: 1100-1106 Tạp chí KH Nông nghiệp VN 2016, tập 14, số 7: 1100-1106 www.vnua.edu.vn CLONING, EXPRESSION AND PURIFICATION OF ABO3 GENE INVOLVED IN DROUGHT STRESS TOLERANCE FROM Arabidopsis thaliana in Escherichia coli Nguyen Xuan Canh*, Nguyen Thi Nhan, Nguyen Thi Phuong Anh, Tran Kim Oanh, Nguyen Thi Thuy Hanh Faculty of Biotechnology, Vietnam National University of Agriculture Email*: nxcanh@vnua.edu.vn Received date: 09.05.2016 Accepted date: 10.08.2016 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. Keywords: Arabidopsis, transcription factor, ABO3, resistance, E. coli, protein expression. Nghiên cứu tách dòng, biểu hiện và tinh