Richael Craig M, Kalyaeva Marina, Chretien Robert C, Yan Hua, Adimulam Sathya, Stivison Artesia, Weeks J Troy, Rommens Caius M
J. R. Simplot Company, Simplot Plant Sciences, Boise, ID, 83706, USA.
Transgenic Res. 2008 Oct;17(5):905-17. doi: 10.1007/s11248-008-9175-6. Epub 2008 Mar 5.
Conventional Agrobacterium-mediated transformation methods rely on complex and genotype-specific tissue culture media for selection, proliferation, and regeneration of genetically modified cells. Resulting transgenic plants may not only contain selectable marker genes but also carry fragments of the vector backbone. Here, we describe a new method for the production of transgenic plants that lack such foreign DNA. This method employs vectors containing the bacterial isopentenyltransferase (ipt) gene as backbone integration marker. Agrobacterium strains carrying the resulting ipt gene-containing "cytokinin" vectors were used to infect explants of various Solanaceous plant species as well as canola (Brassica napus). Upon transfer to hormone-free media, 1.8% to 9.9% of the infected explants produced shoots that contained a marker-free T-DNA while lacking the backbone integration marker. These frequencies often equal or exceed those for backbone-free conventional transformation.
传统的农杆菌介导转化方法依赖于复杂且基因型特异性的组织培养基来进行转基因细胞的筛选、增殖和再生。由此产生的转基因植物不仅可能含有选择标记基因，还可能携带载体骨架片段。在此，我们描述了一种生产不含此类外源DNA的转基因植物的新方法。该方法采用含有细菌异戊烯基转移酶（ipt）基因作为骨架整合标记的载体。携带所得含ipt基因的“细胞分裂素”载体的农杆菌菌株用于感染各种茄科植物以及油菜（甘蓝型油菜）的外植体。转移至无激素培养基后，1.8%至9.9%的受感染外植体产生了不含骨架整合标记且含有无标记T-DNA的芽。这些频率通常等于或超过无骨架传统转化的频率。
