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Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells

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Abstract
A general method is described for the use of tumor-inducing (Ti) plasmids as experimental gene-vectors for plant cells. Intermediate vectors, containing specific Ti plasmid sequences and capable of replication in both E. coli and Agrobacterium strains, were constructed and used for the in vitro introduction of isolated DNA fragments into. predetermined sites of the T-region derived fragments. Site-specific inserts and/or deletions-substitutions inTi plasmids were produced by exchange of the modified T-region sequences for the wild-type sequence by in vivo recombination between intermediate vectors and resident Ti plasmids. This method was applied for the isolation of mutant Ti plasmids affecting either morphogenetic properties or opine synthesis in crown gall tumors. Foreign DNA, inserted at different sites in the T-region of both octopine and nopaline Ti plasmids, was shown to be cotransferred with the T-DNA and to be stably maintained in the transformed plant cells.
Keywords
Ti plasmids, site-specific mutagenesis, plant genetic engineering, Agrobacterium, broad host range cloning vectors

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Chicago
Leemans, Jan, Charles Shaw, Rolf Deblaere, Henri De Greve, Jean-Pierre Hernalsteens, Martine Maes, Marc Van Montagu, and Jeff Schell. 1981. “Site-specific Mutagenesis of Agrobacterium Ti Plasmids and Transfer of Genes to Plant Cells.” Journal of Molecular and Applied Genetics 1: 149–164.
APA
Leemans, Jan, Shaw, C., Deblaere, R., De Greve, H., Hernalsteens, J.-P., Maes, M., Van Montagu, M., et al. (1981). Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells. JOURNAL OF MOLECULAR AND APPLIED GENETICS, 1, 149–164.
Vancouver
1.
Leemans J, Shaw C, Deblaere R, De Greve H, Hernalsteens J-P, Maes M, et al. Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells. JOURNAL OF MOLECULAR AND APPLIED GENETICS. 1981;1:149–64.
MLA
Leemans, Jan, Charles Shaw, Rolf Deblaere, et al. “Site-specific Mutagenesis of Agrobacterium Ti Plasmids and Transfer of Genes to Plant Cells.” JOURNAL OF MOLECULAR AND APPLIED GENETICS 1 (1981): 149–164. Print.
@article{5914466,
  abstract     = {A general method is described for the use of tumor-inducing (Ti) plasmids as experimental gene-vectors for plant cells. Intermediate vectors, containing specific Ti plasmid sequences and capable of replication in both E. coli and Agrobacterium strains, were constructed and used for the in vitro introduction of isolated DNA fragments into. predetermined sites of the T-region derived fragments. Site-specific inserts and/or deletions-substitutions inTi plasmids were produced by exchange of the modified T-region sequences for the wild-type sequence by in vivo recombination between intermediate vectors and resident Ti plasmids. This method was applied for the isolation of mutant Ti plasmids affecting either morphogenetic properties or opine synthesis in crown gall tumors. Foreign DNA, inserted at different sites in the T-region of both octopine and nopaline Ti plasmids, was shown to be cotransferred with the T-DNA and to be stably maintained in the transformed plant cells.},
  author       = {Leemans, Jan and Shaw, Charles and Deblaere, Rolf and De Greve, Henri and Hernalsteens, Jean-Pierre and Maes, Martine and Van Montagu, Marc and Schell, Jeff},
  issn         = {0271-6801},
  journal      = {JOURNAL OF MOLECULAR AND APPLIED GENETICS},
  language     = {eng},
  pages        = {149--164},
  title        = {Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells},
  volume       = {1},
  year         = {1981},
}