转基因玉米的脉冲电泳技术检测及遗传分析

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,对T0代、T1代经除草剂筛选的抗性植株进行PCR检测.结果表明,T1代转基因植株频率明显低于T0代PCR阳性植株率,T0代植株的PCR阳性率与T1代转基因植株频率呈显著正相关(r =0.9874).T1代表现为PCR阳性的植株Southern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,且能从T1代遗传到T2代,外源基因在T2代呈典型的孟德尔分离方式.

Identification and inheritance analysis of transgenic maize by PFGE

The foreign gene(Bar and B.t) was successfully transferred into maize seed by the PFGE(pulsed field gel electrophoresis,PFGE) method.T_0 and T_1 generation plants screened with PPT were further confirmed by PCR.The results revealed that transformation frequency of T_1 generation was much lower than PCR positive frequency of T_0 generation.The both changing frequencies were correlative significant(r=0.9874).T_1 maize plants tested by PCR were confirmed by Southern blot.The results demonstrated that foreign gene was single copy in transformants by PFGE.The exogenous gene inherited from T_1 generation to T_2 generation and showed typical Mendelian segregation in the T_2 generation.