简便实用的转抗除草剂基因后代植株鉴定方法--种子萌发测定法

种子萌发法是检测外源抗性基因在转基因植物中的表达和分析外源抗性基因在转基因植物中遗传行为的简单而直接的方法,以转ｂａｒ基因燕麦后代Ｒ２种子为材料,用不同体积分数的除草剂Ｃｈａｌｌｅｎｇｅ处理种子,萌发的幼苗经ＰＣＲ和Ｓｏｕｔｈｅｒｎ ｄｏｔ ｂｌｏｔ鉴定,发现在０．０３％的除草剂上萌发的幼苗,部分不含ｂａｒ基因,而在０．０７％和０．０９％的除草剂上萌发的幼苗,均含ｂａｒ基因。用０．０７％的除草剂测

Seed Germination Test-A Simple but Effective Way of Analysing Transgenic Progenies Transformed with the bar Gene

It was showed that Seed Germination Test (SGT) was an effective and efficient way of analysing the expression and inheritance of foreign gene in the transgenic oat R3 progeny. The diluted solution of 0.05% to 0.07% commerical herbicide Challenge (150 g/L Glufosinate ammonium, PPT) was found to be optimum in screening the progenies of transgenic cats containing bar gene, produced via microprojectile bombardment. 0.03% of Challenge was too low whereas 0.09% was too high in these experiments. These results were confirmed by the PCR and the Southern dot blot as well as by the segregation ratio analysis. Screening transgenic progenies by the SGT is not only simple and cost-effective, more importantly, it can also obtain transformants with stable highly expressed foreign genes simultaneously.