利用脉冲电泳技术转化玉米获得转基因植株的研究

本研究采用脉冲电泳技术将外源Bar基因和B-t基因转化玉米种胚并直接成苗，通过除草剂抗性初步筛选，再进行PCR检测、Southem杂交验证，探讨了脉冲电泳介导外源基因直接转化玉米种胚的有效性，结果表明：脉冲电泳技术介导玉米转基因是有效可行的，其T0代植株PCR阳性率与T1代植株转化率分别为11．36％和1．04％。在外源基因的转化中，处于不同启动子下的B-t基因和Bar基因共转化频率为90．9％。T1代PCR表现为阳性的植株Southem杂交结果显示脉冲电泳法介导的外源基因在受体中多为单拷贝，且已遗传给后代，并获得稳定表达。

Production of Transgenic Maize Plants by PFGE

In this study, the foreign Bar gene and B.t gene was transferred into maize seeds by the PFGE (pulsed field gel electrophoresis, PFGE) method and the seeds with the foreign gene grew up directly. Through screening the resistance to PPT from seedling, PCR analysis, Southern blot of the resistance plants, the efficiency of exogenous DNA transferring into maize seed was studied. The results are as follow:the technique of transgenesis by PFGE is efficient and feasible. The positive frequency of PCR and transformation is 11.36% and 1.042% respectively. The co-transformation frequency of B.t gene and Bar gene constituted 90.9% in the transformation of exogenous gene. T_1 maize plants tested by PCR were confirmed completely by Southern blot. The results demonstrated that foreign gene was single copy,intergrated and expressed in transformants by PFGE.