Negrotto D, Jolley M, Beer S, Wenck A R, Hansen G
Novartis Agribusiness Biotechnology Research Inc, NABRI, 3054 Cornwallis Road, Research Triangle Park, NC 27709-2257, USA e-mail:
Plant Cell Rep. 2000 Jul;19(8):798-803. doi: 10.1007/s002999900187.
A new selectable marker system has been adapted for use in Agrobacterium-mediated transformation of maize. This selection system utilizes the pmi gene encoding for phosphomannose-isomerase that converts mannose-6-phosphate to fructose-6-phosphate. Only transformed cells are capable of utilizing mannose as a carbon source. Agrobacterium-mediated transformation of immature embryos followed by a pre-selection of 10-14 days prior to selection at a level of 1% mannose and 0.5% sucrose led to the recovery of trangenic lines of a frequency of as high as 30% in about 12 weeks. Molecular and genetic analysis showed that selected plants contained the pmi gene and that the gene was transmitted to the progeny in a Mendelian fashion.
一种新的选择标记系统已被改造用于农杆菌介导的玉米转化。该选择系统利用编码磷酸甘露糖异构酶的pmi基因,该酶可将6-磷酸甘露糖转化为6-磷酸果糖。只有转化细胞能够利用甘露糖作为碳源。农杆菌介导的未成熟胚转化,随后在1%甘露糖和0.5%蔗糖水平下进行10-14天的预筛选,在大约12周内获得了频率高达30%的转基因株系。分子和遗传分析表明,所选植物含有pmi基因,并且该基因以孟德尔方式传递给后代。