番茄基因转化中抗生素筛选压力的确定方法

The Method to Confirm the Selective Concentration of Antibiotic in Genetic Transformation

在以抗生素为筛选标记的基因转化中,确定适宜的筛选压力是提高基因转化率的前提。本研究以卡那霉素为例,通过在10mg/L,20mg/L,30mg/L,50mg/L,100mg/L的筛选压力下,转化和非转化番茄子叶外植体的生长表现、愈伤组织和芽的分化率,来确定适宜的筛选浓度。本实验以该方法找到的适宜的卡那霉素筛选浓度为50mg/L。以此浓度为筛选压力进行遗传转化所获得的部分再生苗经southernblot鉴定,均为转化株。

In order to confirm the optimum selective concentration of antibiotic in genetic transformation, kanamycin was selected as a case in this paper. It had been found that the optimum selective concentration of kanamycin was 50 mg/L by the investigation of the effects upon the growth and differentiation of explants of tomato cotyledon on the media containing different concentrations that were 10 mg/L,20 mg/L,30 mg/L,50 mg/L,100 mg/L, respectively. Some of shoots of transformation that selected by 50 g/L of kanamycin were confirmed by Southern blot.