Abstract
Arabidopsis mutants generated by insertion of the T-DNA from Ti plasmid 3850∶1003 serve as a starting point for the isolation of novel genes. The disrupted plant DNA can be recovered using a plasmid rescue technique utilizing high efficiency electroporation. Rescued plasmids are resistant to ampicillin and contain an origin of replication from pBR322. Plasmids generated from either the left or right border of the T-DNA that carry flanking DNA sequences can be identified by analyzing the products of restriction enzyme digests on agarose gels. The plasmids with flanking sequences can then serve as a starting point for cloning plant sequences that share homology to the DNA at the point of T-DNA insertion.
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Dower, W.J., J.F. Miller and C.W. Ragsdale. 1988. High efficiency transformation ofE. coli by high voltage electroporation. Nucleic Acids Res. 16:6127–6145
Eichholtz, D.A., S.G. Rogers, R.B. Horsch, H.J. Klee, M. Hayford, N.L. Hoffmann, S.B. Bradford, C. Fink, J. Flick, K. O'Connell and R.T. Fraley. 1987. Expression of mouse dihydrofolate reductase gene confers methotrexate resistance in transgenic petunia plants. Somatic Cell Genet. 13:67–76
Errampalli, D., D. Paton, L. Castle, L. Mickelson, K. Hanson, J. Schnall, K. Feldmann and D. Meinke. 1991. Embryonic lethals and T-DNA insertional mutagenesis inArabidopsis. Plant Cell 3:149–157
Feldmann, K.A. and M.D. Marks. 1987.Agrobacterium-mediated transformation of germinating seeds ofArabidopsis thaliana: A non-tissue culture approach. Mol. Gen. Genet. 208:1–9
Feldmann, K.A., M.D. Marks, M.L. Christianson, R.L. Quatrano. 1989. A dwarf mutant ofArabidopsis generated by insertion mutagenesis. Science 243:1351–1354
Feldmann, K.A. 1991: T-DNA insertion mutagenesis inArabidopsis: mutational spectrum. Plant J. 1:71–82
Herman P.L. and M.D. Marks. 1989. Trichome development inArabidopsis thaliana. II. Isolation and complementation of theGLABROUS gene. Plant Cell 1:1051–1055
Marks, M.D. and K.A. Feldmann. 1989. Trichome development inArabidopsis thaliana. I. T-DNA tagging of theGLABROUS1 gene. Plant Cell 1:1043–1050
Medford, J.I., F.J. Behringer, J.D. Callos, K.A. Feldmann. 1992. Normal and abnormal development in theArabidopsis vegetative shoot apex. Plant Cell (in press).
Velten, J. and J. Schell. 1985. Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res. 13:6981–6998
Yanofsky, M.F., H. Ma, J.L. Bowman, G.N. Drews, K.A. Feldmann and E.M. Meyerowitz. 1990. The protein encoded by theArabidopsis homeotic geneagamous resembles transcription factors. Nature 346:35–39
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Behringer, F.J., Medford, J.I. A plasmid rescue technique for the recovery of plant DNA disrupted by T-DNA insertion. Plant Mol Biol Rep 10, 190–198 (1992). https://doi.org/10.1007/BF02668346
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DOI: https://doi.org/10.1007/BF02668346