Designing and construction of appropriate genetic elements for production of glyphosate tolerant transgenic rice

Rice (Oryza sativa L.) is one of the most important food products and the primary source of food for more than a third of the global population. Weed management is an important factor in production of rice in order to achieve high yield and high quality crop. Glyphosate (2-phosphonomethyl-glycine) is a potent non-selective herbicide with a wide range and is widely used in farms. This herbicide inhibits EPSPS, a key enzyme in the shikimate pathway. Development of herbicide-tolerant transgenic plants is the ultimate strategy to reduce the herbicide effect. In this study, the target gene (epsps) that isolated from the Pseudomonas bacteria after screening of soil, it has been shown that this epsps gene has produced an acceptable resistance in bacteria. We needed ideal genetic construct to transfer this gene to plants, so we decided to design this structure. First the epsps gene of the pJET28 vector was transferred using the opportune restriction enzymes into the pBI221 vector. This structure required an appropriate promoter for gene expression into monocotyledonous plants such as rice. We selected the pAct1-D promoter, based on the studies, carried out by the restriction enzymes from the pAct1 carrier to the target vector at the upstream end of the epsps gene. The Intended vector was prepared and transferred into Ecoli.Dh5α bacterial. First the plasmids were extracted from the bacterium, to confirm the cloning of the gene and the promoter in the vector, a polymerase chain reaction was initiated with primers designed for the epsps gene as well as the promoter pAct1-D. Secondly, the presence of a cloned gene and promoter using appropriate restriction enzymes and double digestion enzyme was confirmed. These results of this study can be use in transferring useful and commercially genes to recalcitrant plants.

کلیدواژه ها:

Rice ، Glyphosate Tolerant ، Promoter ، Transformation