pGreen:: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation

被引:1256
作者
Hellens, RP [1 ]
Edwards, EA [1 ]
Leyland, NR [1 ]
Bean, S [1 ]
Mullineaux, PM [1 ]
机构
[1] John Innes Ctr Plant Sci Res, Norwich Res Pk, Norwich NR4 7UH, Norfolk, England
基金
英国生物技术与生命科学研究理事会;
关键词
Agrobacterium; binary vectors; plant transformation; reporter genes; selectable marker genes; Ti vector;
D O I
10.1023/A:1006496308160
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
Binary Ti vectors are the plasmid vectors of choice in Agrobacterium-mediated plant transformation protocols. The pGreen series of binary Ti vectors are configured for ease-of-use and to meet the demands of a wide range of transformation procedures for many plant species. This plasmid system allows any arrangement of selectable marker and reporter gene at the right and left T-DNA borders without compromising the choice of restriction sites for cloning, since the pGreen cloning sites are based on the well-known pBluescript general vector plasmids. Its size and copy number in Escherichia coli offers increased efficiencies in routine in vitro recombination procedures. pGreen can replicate in Agrobacterium only if another plasmid, pSoup, is co-resident in the same strain. pSoup provides replication functions in trans for pGreen. The removal of RepA and Mob functions has enabled the size of pGreen to be kept to a minimum. Versions of pGreen have been used to transform several plant species with the same efficiencies as other binary Ti vectors. Information on the pGreen plasmid system is supplemented by an Internet site (http://www.pgreen.ac.uk) through which comprehensive information, protocols, order forms and lists of different pGreen marker gene permutations can be found.
引用
收藏
页码:819 / 832
页数:14
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