McBride K E, Summerfelt K R
Calgene Inc., Davis, CA 95616.
Plant Mol Biol. 1990 Feb;14(2):269-76. doi: 10.1007/BF00018567.
Improved plant transformation vectors were constructed which utilize the pRiHRI origin of replication for highly stable maintenance in Agrobacterium tumefaciens, the ColE1 origin of replication for high copy maintenance in Escherichia coli, and a gentamycin resistance gene as a strong selectable marker for bacteria. Concise T-DNA elements were engineered with border sequences from the TL-DNA of pTiA6, the Tn5 neomycin phosphotransferase gene (npt II) expressed from either CaMV 35S or mannopine synthase (mas) promoters, and the lac Z' gene segment from pUC18 as a source of unique restriction sites as well as an insertional inactivation marker for cloned DNA. The order of T-DNA components in all vectors is left border, plant marker cassette, lac Z', and right border, respectively. The prototype vector, pCGN1547, was shown to be very stable in A. tumefaciens strain LBA4404 and to act as an efficient donor of T-DNA in tomato transformation experiments. Use of the other vectors is also described.
构建了改良的植物转化载体，其利用发根农杆菌RiHRI复制起点在根癌农杆菌中进行高度稳定的维持，利用ColE1复制起点在大肠杆菌中进行高拷贝维持，并利用庆大霉素抗性基因作为细菌的强选择标记。构建了简洁的T-DNA元件，其具有来自pTiA6的TL-DNA的边界序列、由花椰菜花叶病毒35S启动子或甘露碱合成酶（mas）启动子表达的Tn5新霉素磷酸转移酶基因（npt II），以及来自pUC18的lac Z'基因片段，作为独特限制酶切位点的来源以及克隆DNA的插入失活标记。所有载体中T-DNA组件的顺序分别为左边界、植物标记盒、lac Z'和右边界。原型载体pCGN1547在根癌农杆菌菌株LBA4404中显示出非常稳定，并在番茄转化实验中作为T-DNA的有效供体。还描述了其他载体的使用情况。
