Design and make up 35S::DREB1A molecular construct to produce environmental stress tolerant plants

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Environmental stresses affect plant growth and cause losses worth hundreds of million dollars of agricultural industry each year. Many genes induced in response to environmental stresses. The DREB1A gene is a stress-inducible transcription factor that its overexpression increased tolerance to environmental stress in transgenic plants. DNA was extracted from Arabidopsis thaliana var. Col.0 plants and DREB1A gene amplified by particular primers. After purification, PCR products were cloned into pGEMT-EASY vector and transferred into E. coli strain DH5α competent cells. Blue/white colonies were analyzed and present of the DREB1A gene revealed by restriction analysis and sequencing test. A 668-bp XbaI/BamHI digested fragment of DREB1A gene from pGEMT::DREB1A construct cloned in pBI121 plasmid. The recombinant plasmids were transferred into Agrobacterium tumefaciens cells strain of LBA4404 and transformed cells were cultured on selective medium that supplied with kanamycin and rifampycin. As a result of gel electrophoresis of Agrobacterium colonies-PCR product, band existing on expected size show that the DREB1A gene was cloned in to pBI121 binary vector. At time, Agrobacterium cells containing of pBI121::DREB1A construct are using for product of environmental stress tolerant plants.