Zhu Y J, Agbayani R, Moore P H
Hawaii Agriculture Research Center, 99-193 Aiea Heights Drive, Aiea, Hawaii 96701, USA.
Plant Cell Rep. 2004 Apr;22(9):660-7. doi: 10.1007/s00299-004-0755-5. Epub 2004 Jan 29.
Chemical-based selection for plant transformation is associated with a number of real and perceived problems that might be avoided through visual selection. We have used green fluorescent protein (GFP), as a visual selectable marker to produce transformed papaya ( Carica papaya) plants following microprojectile bombardment of embryogenic callus. GFP selection reduced the selection time from 3 months on a geneticin (G418) antibiotic-containing medium to 3-4 weeks. Moreover, GFP selection increased the number of transformed papaya plants by five-to eightfold compared to selection in the presence of antibiotics. Overall, the use of GFP for selecting transgenic papaya lines improved our throughput for transformation by 15- to 24-fold while avoiding the drawbacks associated with the use of antibiotic resistance-based selection markers.
基于化学物质的植物转化筛选存在许多实际和认知上的问题，而通过视觉筛选或许可以避免这些问题。我们利用绿色荧光蛋白（GFP）作为视觉选择标记，在对胚性愈伤组织进行微粒轰击后培育出了转基因番木瓜（番木瓜属）植株。GFP筛选将筛选时间从在含遗传霉素（G418）抗生素的培养基上的3个月缩短至3 - 4周。此外，与在抗生素存在下进行筛选相比，GFP筛选使转基因番木瓜植株的数量增加了五至八倍。总体而言，使用GFP筛选转基因番木瓜品系将我们的转化通量提高了15至24倍，同时避免了与使用基于抗生素抗性的选择标记相关的缺点。
