Nucleic Acids Research, 1985, Vol. 13, No. 13 4777-4788
© 1985
Articles |
Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants
Laboratorium voor Genetische Virologie, Vrije Universiteit Brussel B-1640 St.-Genesius-Rode, Belgium
Received March 28, 1985. Revised June 14, 1985. Accepted June 14, 1985.
A two-component cloning system to transfer foreign DNA into plants was derived from the octopine Ti plasmid pTiB6S3. pGV2260 is a non-oncogenic Ti plasmid from which the T-region is deleted and substituted by pBR322. pGVB31 is a streptomycin-resistant pBR325 derivative that contains a kanamycin resistance marker gene for plant cells and a site for cloning foreign genes between the 25-bp border sequences of the octopine T-region. Conjugative transfer of pGV831 derivatives to Agrobacterium and cointegration by homologous recombination between the pBR322 sequences present on pGV831 and pGV2260, can be obtained in a single step. Strains carrying the resulting cointegrated plasmids transfer and integrate T-DNA into the genome of tobacco protoplasts, and transformed tobacco calli are readily selected as resistant to kanamycin. Intact plants containing the entire DNA region between the T-DNA borders have been regenerated from such clones. In view of these properties we present pGV831 and its derivatives as vectors for efficient integration of foreign genes into plants.
*Present address: Plant Genetic Systems, Plateaustraat 22, B-9000 Gent, Belgium
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