转基因玉米的脉冲电泳技术检测及遗传分析

Identification and inheritance analysis of transgenic maize by PFGE

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。

The foreign gene （Bar and B. t） was successfully transferred into maize seed by the PFGE （pulsed field gel electrophoresis, PFGE） method. T0 and T1 generation plants screened with PPT were further confirmed by PCR. The results revealed that transformation frequeney of T1 generation was much lower than PCR positive frequency of To generation. The both changing frequencies were correlative significant （ r = 0. 9874）. T1 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 T1 generation to T2 generation and showed typical Mendelian segregation in the T2 generation.