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PMID: 16453603 Published · ppublish English Journal Article

New cloning vehicles for transformation of higher plants.

The EMBO journal ·Vol. 4 ·No. 2 ·1985-02-00 ·Pages 277-84

An G, Watson BD, Stachel S, Gordon MP, Nester EW

Abstract

We have constructed a set of small vectors based on the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens which allow the transfer of exogenous DNA into plant chromosomes. These vectors contain: (i) a chimeric gene containing the transcriptional control signals from the nopaline synthase gene and the coding sequence for neomycin phosphotransferase; (ii) the ColE1 replicon; (iii) the cos site of bacteriophage lambda; (iv) the border sequences from the ends of the T-DNA region of the Ti plasmid; and (v) a wide host range replicon. Due to the small size of these cosmid vectors, DNA fragments up to 35 kbp can be inserted by an in vitro packaging method in Escherichia coli. The ability of these vectors to be stably replicated in both E. coli and A. tumefaciens allows their subsequent transfer to and maintenance in Agrobacterium without intermediate genetic manipulations. We demonstrate that DNA cloned into these vectors in A. tumefaciens can efficiently transform plants when in trans with a wild-type Ti plasmid which donates the functions necessary for DNA transfer and integration. We also show that only the right border of the T-DNA is necessary for DNA transformation.

Authors & Affiliations
5 authors, click to expand affiliations / ORCID
An G
Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, USA.
Watson B D
Stachel S
Gordon M P
Nester E W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1985-02-00
Pages
277-84
Language
English
Region
England
NLM ID
8208664
PMCID
PMC554183
External Links
PubMed source PMC Full Text View journal details
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