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New cloning vehicles for transformation of higher plants
Author(s) -
An G.,
Watson B.D.,
Stachel S.,
Gordon M.P.,
Nester E.W.
Publication year - 1985
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1985.tb03626.x
Subject(s) - biology , agrobacterium tumefaciens , replicon , plasmid , transfer dna , cosmid , t dna binary system , ti plasmid , transformation (genetics) , multiple cloning site , in vitro recombination , microbiology and biotechnology , agrobacterium , genetics , dna , cloning vector , molecular cloning , gene , expression vector , vector (molecular biology) , complementary dna , recombinant dna
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 λ; (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.