Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants
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Progress in plant molecular biology has depended heavily on the availability of effective vectors for plant cell transformation and heterologous expression. In this paper we describe the structures of a wide array of plasmids which have proved extremely effective in (a) plant transformation, (b) expression of heterologous genes and (c) assaying the activity of transposons in transgenic plants. Constructs that confer resistance to kanamycin, hygromycin, streptomycin, spectinomycin and phosphinotricin, or that confer β-glucuronidase (GUS) gene expression are presented. Binary vector constructs that carry polylinkers of the pUC and Bluescript types are also described. Plasmids that permit the expression of any heterologous reading frame from either nopaline synthase (nos) or octopine synthase (ocs) promoters, as well as the cauliflower mosaic virus 35S promoter, using either the nopaline synthase or octopine synthase 3′ polyadenylation sequences, are presented. These constructs permit a choice of orientation of the resulting transgene of interest, relative to the orientation of the selection marker gene. Most of the plasmids described here are publicly available.
Keywordsbinary vector plant expression plasmid kanamycin resistance spectinomycin resistance hygromycin resistance phosphinotricin resistance streptomycin resistance β-glucuronidase
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- Chinault, A. C., Blakesley, V. A., Roessler, E., Willis, D. G., Smith, C. A., Cook, R. G. and Fenwick, R. G. (1986) Characterization of transferable plasmids fromShigella flexneri 2a that confer resistance to trimethoprim, streptomycin and sulfonamides.Plasmid 15, 119–31.PubMedCrossRefGoogle Scholar
- DeBlock, M., Botterman, J., Vandewiele, M., Dockx, J., Thoen, C., Gossele, V., RaoMovva, N., Thompson, C., Van Montagu, M. and Leemans, J. (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme.EMBO J. 6, 2513–8.Google Scholar
- DeGreve, H., Dhaese, P., Seurinck, J., Lemmers, S., van Montagu, M. and Schell, J. (1983) Nucleotide sequence and transcript map of theAgrobacterium tumefaciens Ti plasmid-encoded octopine synthase gene.J. Mol. Appl. Genet. 1, 499–511.Google Scholar
- Harpster, M. H., Townsend, J. A., Jones, J. D. G., Bedbrook, J. and Dunsmuir, P. (1988) Relative strengths of the 35S cauliflower mosaic virus, 1′, 2′, and nopaline synthase promoters in transformed tobacco, sugarbeet and oilseed rape callus tissueMol. Gen. Genet. 212, 182–90.PubMedCrossRefGoogle Scholar
- Huffman, G. A., Beach, L. R., Martich, J. M., Fall, M. M., Sims, L. E., Burrus, M., Marsh, W. A., Maddock, S. E. and Bauer, R. E. (1991) Optimizing plant expression vectors.J. Cell Biochem. Suppl.15A, 98.Google Scholar
- Jones, J. D. G., Harper, L., Carland, F., Ralston, E. and Dooner, H. (1991) Reversion and altered variegation of an SPT::Ac allele in tobacco.Maydica 36, 329–35.Google Scholar
- Rogers, S. G., Klee, H. J., Horsch, R. B. and Fraley, R. T. (1987) Improved vectors for plant transformation: expression cassette vectors and new selectable markers.Methods in Enzymol. 153, 253–305.Google Scholar
- Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989)Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
- White, J., Chang, S-Y. P., Bibb, M. J. and Bibb, M. J. (1991) A cassette containing thebar gene ofStreptomyces hygroscopicus: a selectable marker for plant transformation.Nucl. Acids Res. 18, 1062.Google Scholar