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Cloning and Characterization of Alcohol Dehydrogenase (Adh) Promoter Region for Expression Under Submergence and Salinity Stress

机译:淹没和盐分胁迫下表达的酒精脱氢酶(Adh)启动子区的克隆和鉴定

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The characterization of promoter is important for developing stress tolerant crops as well as understanding the role of promoters in regulating gene expression. The current study was initiated with an aim to characterize the Adh promoter under salinity and submergence stress in rice calli. The upstream regions (~1kb) of the Adh gene was amplified from the genomic DNA of Arabidopsis (Columbia Ecotype). The amplified product was then cloned successively into an entry and promoter-characterization binary destination vector having the reporter gene β-glucuronidase (GUS) by applying Gateway Technology. A positive clone was confirmed by applying PCR, restriction digestion and sequencing. The construct was then transformed into Agrobacterium tumefaciens LBA4404 strain and rice calli infected with the latter. In both salt and submergence stresses, Adh could selectively express GUS gene activity up to two-fold compared to control. Plant Tissue Cult. & Biotech. 24(1): 111-120, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19252
机译:启动子的表征对于开发耐逆性作物以及理解启动子在调控基因表达中的作用很重要。当前的研究是为了表征水稻愈伤组织在盐度和淹没胁迫下的Adh启动子而进行的。从拟南芥的基因组DNA(哥伦比亚生态型)扩增Adh基因的上游区域(〜1kb)。然后通过应用Gateway Technology将扩增的产物连续克隆到具有报告基因β-葡糖醛酸糖苷酶(GUS)的进入和启动子表征的二进制目的载体中。通过应用PCR,限制性酶切和测序确认阳性克隆。然后将构建体转化到根癌农杆菌LBA4404菌株中,并用后者感染水稻愈伤组织。在盐胁迫和淹没胁迫下,Adh可以选择性表达GUS基因活性,是对照组的两倍。植物组织崇拜。和生物技术。 24(1):111-120,2014(6月)D.O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19252

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