Plant Molecular Biology

, Volume 20, Issue 6, pp 1203–1207

A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome

  • Andrew P. Gleave
Update Section Short Communication

DOI: 10.1007/BF00028910

Cite this article as:
Gleave, A.P. Plant Mol Biol (1992) 20: 1203. doi:10.1007/BF00028910
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Abstract

A versatile gene expression cartridge and binary vector system was constructed for use in Agrobacterium-mediated plant transformation. The expression cartridge of the primary cloning vector, pART7, comprises of cauliflower mosaic virus Cabb B-JI isolate 35S promoter, a multiple cloning site and the transcriptional termination region of the octopine synthase gene. The entire cartridge can be removed from pART7 as a Not I fragment and introduced directly into the binary vector, pART27, recombinants being selected by blue/white screening for β-galactosidase. pART27 carries the RK2 minimal replicon for maintenance in Agrobacterium, the ColE1 origin of replication for high-copy maintenance in Escherichia coli and the Tn7 spectinomycin/streptomycin resistance gene as a bacterial selectable marker. The organisational structure of the T-DNA of pART27 has been constructed taking into account the right to left border, 5′ to 3′ model of T-DNA transfer. The T-DNA carries the chimaeric kanamycin resistance gene (nopaline synthase promoter-neomycin phosphotransferase-nopaline synthase terminator) distal to the right border relative to the lacZ′ region. Utilisation of these vectors in Agrobacterium-mediated transformation of tobacco demonstrated efficient T-DNA transfer to the plant genome.

Key words

Agrobacterium binary vector CaMV 35S gene expression β-glucuronidase Nicotiana plumbaginifolia 

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Andrew P. Gleave
    • 1
  1. 1.Molecular Genetics Group, Plant Improvement DivisionHorticulture and Food Research Institute of New Zealand Ltd.AucklandNew Zealand

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