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实验构建热稳定性聚合酶的基因表达双元载体,利用花粉介导法将热稳定性DNA聚合酶基因导入玉米幼胚,获得耐热转基因玉米株系。用热稳定性DNA聚合酶基因构建表达载体;然后利用花粉介导法进行转化,花粉收集自仲恺自交系玉米甜1号和自交系糯2号,将花粉与构建好的基因表达载体混合,利用花粉介导法进行转化;将收集到的T1代种子用PCR凝胶电泳技术和Southern杂交技术进行检测,选出阳性的转化株;在T2代经过田间耐热压力试验结合测定的玉米生理指标,筛选出9个具有良好耐热性玉米株系。
The binary vector for gene expression of thermostable polymerase gene was constructed experimentally. The gene of thermostable DNA polymerase was introduced into immature embryos of maize by pollen-mediated method to obtain the heat-resistant transgenic maize. The expression vector was constructed with the thermostable DNA polymerase gene. The pollen was then transformed by pollen-mediated method. Pollen was collected from Zhongkai inbred line corn sweet No.1 and inbred line glutinous 2, and the pollen was mixed with the constructed gene expression vector , And transformed by pollen mediated method. The collected T1 generation seeds were detected by PCR gel electrophoresis and Southern hybridization, and the positive transformants were selected. In the T2 generation, after the field heat stress test and the corn physiological Indicators, screened nine have a good heat-resistant corn lines.