抗草甘膦EPSPS转基因棉花的研究

林敏教授课题组从具有草甘膦抗性的荧光假单胞菌G2中克隆了EPSP合酶基因aroAG2，根据aroAG2的氨基酸序列，按照双子叶植物偏爱密码子人工合成aroAG2M基因。我们利用aroAG2M基因和Impactvector1.4，构建了以菊花Rubisco启动子和叶绿体信号肽驱动aroAG2M基因高效表达的植物表达载体Pim1.4-epsps-plus，以农杆菌介导法转化烟草，转基因烟草经PCR和Southern检测，证明外源基因已整合到基因组中，转基因烟草具有草甘膦抗性。以花粉管通道法转化棉花，经草甘膦抗性筛选得到T1代转基因棉花3株，转基因棉花的PCR和Southern检测结果均为阳性，说明目的基因已通过花粉管通道法导入棉花。

The Study of Glyphosate-resistance Transgenic Cotton

Dr lin who works in the biotechnology research institute has isolated and cloned the structural gene aroAG2 coding for EPSP Synthase sequence from Pseudomonas fluorescens G2. The aroAG2M gene was artificially synthesized according to codon usage preferred by dicotyledonous plants.Impactvector1.4 with the chloroplast signal peptide is one of high performing vectors for expression in plant. The aroAG2M gene was placed under the Rubisco promoter and then a plant expression vector was constructed. Transgenic tobacco were obtained by Agrobacterium-mediated transformation．PCR and Southern analysis of transgenic tabacco plants indicated that the exogenous genes were integrated into the genomes of the transgenic plants,and in transgenic plants herbicide-resistance to glyphosate reaches 4‰.The vector was used for cotton transformation by Pollen Tube Pathway Transgenic Technique. PCR and Southern analysis of transgenic cotton plants showed the target gene was integrated into the genome. To date, T1 generation of transgenic plants have been obtained. Transgenic plants are resistant to glyphosate herbicide in field．