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针对草甘膦结合位点,对高粱5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)基因进行4种定点修饰,将修饰后的基因分别导入到玉米中。通过对转化体的抗性鉴定,确定将高粱EPSPS基因106位的脯氨酸变为丝氨酸(P106S)能够赋予转基因玉米草甘膦抗性。在喷施4倍的草甘膦时抗性事件CL38-1不产生药害。Southern杂交结果表明,目标基因在该转化事件中稳定遗传,转化其余3种EPSPS基因的植株对草甘膦没有足够的抗性。
According to the glyphosate binding sites, four kinds of site-directed modifications were carried out on the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene of sorghum. The modified genes were introduced into maize separately. Through the identification of transformants, it was determined that the transformation of proline at position 106 of sorghum EPSPS to serine (P106S) conferred glyphosate resistance in transgenic maize. Resistance event CL38-1 did not produce phytotoxicity when 4 times glyphosate was sprayed. Southern hybridization results showed that the target gene was stably inherited in the transformation event and the plants transformed with the other three EPSPS genes did not have sufficient resistance to glyphosate.