IMPROVED BINARY VECTORS FOR AGROBACTERIUM-MEDIATED PLANT TRANSFORMATION

被引:298
作者
MCBRIDE, KE
SUMMERFELT, KR
机构
[1] Calgene Inc., Davis, 95616, CA
关键词
Agrobacterium; binary vectors; ColE1; replicon; lac Z′; plant transformation; pRi replicon;
D O I
10.1007/BF00018567
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. © 1990 Kluwer Academic Publishers.
引用
收藏
页码:269 / 276
页数:8
相关论文
共 24 条