Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants.
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. pGV831 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.
D001120:Arginine / Q000031:analogs & derivatives* / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005822:Genetic Vectors; D009693:Nucleic Acid Hybridization; D010944:Plants / Q000235:genetics*; D010947:Plants, Toxic; D010957:Plasmids; D011523:Protoplasts; D012231:Rhizobium / Q000235:genetics*; D014026:Nicotiana; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA; C005394:octopine; D001120:Arginine