Abstract
A minimal T-DNA binary vector was used for Agrobacterium-mediated transfer of a chimeric T4 lysozyme gene located next to the left border, and transgenic potato plants which expressed T4 lysozyme protein were identified and further analysed. Frequent rearrangements of T4 lysozyme transgenes were detected. A vector derivative containing two matrix associated regions (MARs) flanking its multiple cloning site was constructed. In transgenic potato plants, reduced variability in gene expression due to position effects was detected. When either the donor vector contained MAR sequences, or when vector pPCV701 which contains a pBR322 fragment next to the left border were used, only relatively few rearrangements were observed. However, when the T4 lysozyme gene was driven by a CaMV 35S promoter modified by multiplied enhancer region carrying either 2 or 4 elements, frequent rearrangements were again obtained.
References
Breyne P, Van Montagu M, Depicker A, Gheysen G: Characterization of a plant scaffold attachment region in a DNA fragment that normalizes transgene expression in tobacco. Plant Cell 4: 463–471 (1992).
Düring K: A plant transformation vector with a minimal T-DNA. Transgen Res 3: 138–140 (1994).
Düring K, Porsch P, Fladung M, Lörz H: Transgenic potato plants resistant to the phytopathogenic bacterium Erwinia carotovora. Plant J 3: 587–598 (1993).
Gheysen G, Villarroel R, Van Montagu M: Illegitimate recombination as a model for T-DNA integration. Genes Dev 5: 287–297 (1991).
Hayashi H, Czaja I, Lubenow L, Schell J, Walden R: Activation of a plant gene by T-DNA tagging: auxinindependent growth in vitro. Science 258: 1350–1353 (1992).
Koncz C, Olsson O, Langridge WHR, Schell J: Expression and assembly of functional bacterial luciferase in plants. Proc Natl Acad Sci USA 84: 131–135 (1987).
Mayerhofer R, Koncz-Kalman Z, Nawrath C, Bakkeren G, Crameri A, Angelis K, Redei GP, Schell J, Hohn B, Koncz C: T-DNA integration: a mode of illegitimate recombination in plants. EMBO J 10: 697–704 (1991).
Odell JT, Nagy F, Chua NH: Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313: 810–812 (1985).
Van Engelen FA, Schouten A, Molthoff JW, Roosien J, Salinas J, Dirkse WG, Schots A, Bakker J, Gommers FJ, Jongsma MA, Bosch D, Stiekema WJ: Coordinate expression of antibody subunit genes yields high levels of functional antibodies in roots of transgenic tobacco. Plant Mol Biol 26: 1701–1710 (1994).
Velten J, Schell J: Selectionexpression plasmid vectors for use in genetic transformation of higher plants. Nucl Acids Res 13: 6981–6998 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Porsch, P., Jahnke, A. & Düring, K. A plant transformation vector with a minimal T-DNA II. Irregular integration patterns of the T-DNA in the plant genome. Plant Mol Biol 37, 581–585 (1998). https://doi.org/10.1023/A:1006035500546
Issue Date:
DOI: https://doi.org/10.1023/A:1006035500546