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

Date Received: 09-05-2016

Date Published: 10-08-2016

##submissions.doi##: https://doi.org/10.31817/tckhnnvn.2016.14.7.

Views

0

Downloads

0

How to Cite:

Canh, N., Nhan, N., Anh, N., Oanh, T., & Hanh, N. (2016). CLONING, EXPRESSION AND PURIFICATION OF ABO3 GENE INVOLVED IN DROUGHT STRESS TOLERANCE FROM Arabidopsis thaliana in Escherichia coli. Vietnam Journal of Agricultural Sciences, 14(7), 1100–1106. https://doi.org/10.31817/tckhnnvn.2016.14.7.

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

Nguyen Xuan Canh (*) 1 , Nguyen Thi Nhan 1 , Nguyen Thi Phuong Anh 1 , Tran Kim Oanh 1 , Nguyen Thi Thuy Hanh 1

  • Tác giả liên hệ: [email protected]
  • 1 Faculty of Biotechnology, Vietnam National University of Agriculture

    • Keywords

    Arabidopsis, transcription factor, ABO3, resistance, E. coli, protein expression

    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.

    References

    Ingram J and Bartels D. (1996). The molecular basis of dehydration tolerance in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol., 47: 377-403.

    Olivier V. A, Botao Z, Simon L, Reena N, and James W. (2013). AtWRKY40 and AtWRKY63 Modulate the Expression of Stress-Responsive Nuclear Genes Encoding Mitochondrial and Chloroplast Proteins. Plant Physiol., 162: 254-271.

    Price A. H, Cairns J. E, Horton P, Jones H. G, and Griffiths H. (2002). Linking drought-resistance mechanisms to drought avoidance in upland rice using a QTL approach: progress and new opportunities to integrate stomatal and mesophyll responses. J. Exp. Bot., 53: 989-1004.

    Ramankutty N., Evan A. T, Monfreda C, and Foley J. A. (2008). Farming the planet. Part 1: the geographic distribution of global agricultural lands in the year 2000 Glob. Biogeochem. Cycles, 22 GB1003.

    Ren X, Chen Z, Liu Y, Zhang H, Zhang M, Liu Q, Hong X, Zhu J. K, and Gong Z. (2010). ABO3, a WRKY transcription factor, mediates plant responses to abscisic acid and drought tolerance in Arabidopsis. Plant J., 63: 417-429.

    Rosegrant M. W and Cline S. A. (2003). Global food security: challenges and policies. Science, 302: 1917-1919.

    Siebert S, Döll P, Hoogeveen J, Faures J. M, Frenken K, and Feick S. (2005). Development and validation of the global map of irrigation areas. Hydrol. Earth Syst. Sci., 9: 535-47. doi:10.5194/hess-9-535-2005.

    Yancey P. H, Clark M. E, Hand S. C, Bowlus R. D, and Somero G. N.(1982). Living with water stress: evolution of osmolyte systems. Science, 217: 1214-1222.