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

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...

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Bibliographic Details
Published in:Plant molecular biology 1992-12, Vol.20 (6), p.1203-1207
Main Author: Gleave, A.P
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Agrobacterium tumefaciens - genetics
Base Sequence
beta-glucuronidase
Biological and medical sciences
Biotechnology
Cauliflower mosaic virus
drug resistance
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
genetic transformation
Genetic Vectors
genetically modified organisms
kanamycin
Methods. Procedures. Technologies
Molecular Sequence Data
Nicotiana plumbaginifolia
Oligodeoxyribonucleotides - chemistry
Plants - genetics
Plasmids
promoter regions
recombinant DNA
reporter genes
streptomycin
Transformation, Genetic
vectors
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary: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 beta-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.
ISSN:0167-4412
1573-5028
DOI:10.1007/BF00028910