Summary
DNA from a bacterial plasmid containing the T-DNA border sequences of Agrobacterium tumefaciens was transferred into the nucleus or the cytoplasm of tobacco mesophyll protoplasts by microinjection. Following culture in hanging drops, some of these protoplasts produced calli containing the foreign DNA sequences. Evidence for the presence of the injected plasmid DNA in these calli was provided by Southern hybridization analysis. The results demonstrated that random portions of the bacterial plasmid were linked to plant DNA and that integration did not occur at the T-DNA borders present on the injected plasmid. The average number of integrated copies ranged from less than one to 1–2 per tobacco genome. The frequency of integration averaged 14% with intranuclear injections compared to 6% with cytoplasmic injections. With further refinement, the use of microinjection may allow the introduction of many different types of genetic elements into plants.
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Communicated by R.B. Goldberg
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Crossway, A., Oakes, J.V., Irvine, J.M. et al. Integration of foreign DNA following microinjection of tobacco mesophyll protoplasts. Mol Gen Genet 202, 179–185 (1986). https://doi.org/10.1007/BF00331634
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DOI: https://doi.org/10.1007/BF00331634