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利用PCR技术扩增含有hrpZpsta基因的克隆载体pMD18-T-hrpZpsta,以植物表达载体pBI121为基础,将PCR产物插入到此植物表达载体中。利用农杆菌介导的大豆子叶节转化法将构建好的表达载体导入大豆品种吉农28中,选用筛选条件为100mg·L-1的卡那霉素选择培养基培养,对获得的转基因植株进行PCR检测,结果从部分抗性植株中扩增出hrpZpsta基因,初步证明hrpZpsta基因成功转入到受体大豆中,获得了转基因阳性植株。
The cloning vector pMD18-T-hrpZpsta containing hrpZpsta gene was amplified by PCR, and the PCR product was inserted into the plant expression vector based on the plant expression vector pBI121. The Agrobacterium-mediated transformation of soybean cotyledons section was used to introduce the constructed expression vector into the soybean variety Jinong 28, and the selected transgenic plants were selected with kanamycin selection medium with a screening condition of 100 mg · L -1, and the obtained transgenic plants were subjected to PCR results showed that the hrpZpsta gene was amplified from the partially resistant plants. It was initially proved that the hrpZpsta gene was successfully transferred into the recipient soybean and transgenic plants were obtained.