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Targeted Insertion of Foreign Genes into the Tobacco Plastid Genome without Physical Linkage to the Selectable Marker Gene

Bio/Technology volume 13, pages 791–794 (1995)Cite this article

Abstract

To determine whether targeted DNA insertion into the tobacco plastid genome can be obtained without physical linkage to a selectable marker gene, we carried out biolistic transformation of chloroplasts in tobacco leaf segments with a 1:1 mix of two independently targeted antibiotic resistance genes. Plastid transformants were selected by spectinomycin resistance due to expression of an integrated aadA gene. Integration of the unselected kanamycin resistance (kan) gene into the same plastid genome was established by Southern probing in ∼20% of the spectinomycin-selected clones. Efficient cotransformation will facilitate targeted plastid genome modification without physical linkage to a marker gene.

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Author notes

  1. Helaine Carrer

    Present address: Centro de Biotecnologia Agricola, Departamento de Quimica, ESA “Luiz de Queiroz”, USP, Piracicaba-SP, 13418-900, Brazil

Authors and Affiliations

  1. Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, NJ, 08855-0759

    Pal Maliga

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  1. Helaine Carrer
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  2. Pal Maliga
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Correspondence to Pal Maliga.

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Carrer, H., Maliga, P. Targeted Insertion of Foreign Genes into the Tobacco Plastid Genome without Physical Linkage to the Selectable Marker Gene. Nat Biotechnol 13, 791–794 (1995). https://doi.org/10.1038/nbt0895-791

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  • DOI: https://doi.org/10.1038/nbt0895-791

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