Zhao Fu-Yong, Li Yun-Feng, Xu Peilin
State Key Laboratory of Biocontrol, Biotechnology Research Center, Zhongshan University, Guangzhou, 510275, The People's Republic of China.
Biotechnol Lett. 2006 Aug;28(15):1199-207. doi: 10.1007/s10529-006-9078-7. Epub 2006 Jun 24.
The most economically significant Chinese cotton cultivar (Gossypium hirsutum L. cv. Zhongmian 35) was transformed via Agrobacterium tumefaciens-mediated DNA transfer. The aroA-M1 gene that confers resistance to the glyphosate was fused with a chloroplast-transit peptide of Arabidopsis thaliana 5-enolpyruvyl-3-phosphoshikimate synthase (ASP) and expressed in cotton plants under the control of a CaMV35S promoter. Transgenic plants were directly selected on medium containing glyphosate. Thirty-four independent transgenic lines were obtained after selection, giving a maximal 1.9% transformation frequency. The integration and expression of the aroA-M1 gene in T(0) plants and T(1) progeny were confirmed using DNA hybridization, Western blot and PCR techniques. An increased resistance of T(0) and T(1 )transgenic plants towards glyphosate was also observed.
通过根癌农杆菌介导的DNA转移，对中国经济意义最为重大的棉花品种（陆地棉品种中棉35）进行了转化。赋予草甘膦抗性的aroA - M1基因与拟南芥5 - 烯醇丙酮酸 - 3 - 磷酸莽草酸合酶（ASP）的叶绿体转运肽融合，并在花椰菜花叶病毒35S启动子的控制下在棉花植株中表达。在含有草甘膦的培养基上直接筛选转基因植株。筛选后获得了34个独立的转基因株系，最高转化频率为1.9%。使用DNA杂交、蛋白质免疫印迹和PCR技术证实了aroA - M1基因在T(0)植株和T(1)后代中的整合与表达。还观察到T(0)和T(1)转基因植株对草甘膦的抗性增强。
