GFP绿色荧光法辅助筛选玉米转基因植株

利用GFP(Green Fluorescence Protein)绿色荧光蛋白基因作为辅助性筛选标记基因,基因枪法轰击玉米自交系A188和Qi319未成熟胚的胚性愈伤组织。瞬间表达实验结果表明,轰击后GFP表达可持续近20 d,轰击后10 d左右表达量仍保持在较高的水平。采用潮霉素筛选、GFP荧光法鉴定,获得一批玉米转基因植株,转化频率为0.33%~0.47%。分子鉴定和后代分析的结果表明,外源基因已经整合到了玉米基因组中,并能够稳定地表达和遗传。GFP作为辅助性筛选标记基因可以直观、快速和有效地消除玉米遗传转化工作中的假阳性现象。

Production of Transgenic Maize Plants via the GFP Green Fluorescence-assisted Selection

Using a plasmid containing GFP(Green Fluorescence Protein) gene as the visual marker, thecalli of immature embryos of maize inbred lines A188 and Qi319 were bombarded. In a transient expression experiment, the GFP fluorescence was observed within 20 days and kept at a high level in 10 days after bombardment. Under the selection of hygromycin and by the visual check of GFP fluorescence, a few transgenic maize plants were produced with a frequency of 0.33%-0.47%. The identity of the transgenics was confirmed by the molecular analysis assays and genetic analysis experiments of offspring later. This work suggests that GFP is a good visual marker to pick up the real transformants from the resistant calli and shoots of maize.