Transgene Organisation in Potato After Particle Bombardment-mediated (co-)Transformation Using Plasmids and Gene Cassettes View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2003-08

AUTHORS

Andrea Romano, Krit Raemakers, Jamila Bernardi, Richard Visser, Hans Mooibroek

ABSTRACT

Protocols for efficient co-transformation of potato internodes with genes contained in separate plasmids or gene cassettes (i.e., linear PCR fragments comprising a promoter-gene-terminator) using particle bombardment were established. Twenty-eight out of 62 (45%) and 11 out of 65 (17%) plants transformed with a plasmid containing the selectable marker contained one and two additional non-selected genes, respectively. When gene cassettes were used in transformation, six out of eight plants were co-transformed. Expression analysis showed that 75–80% of the plants transformed with two transgenes expressed both of them, irrespective of the use of plasmids or gene cassettes. Thirty-eight plants containing the gusA reporter-gene and the nptII selectable-marker have been characterised with respect to the molecular organisation of the donor DNAs. Seventeen out of 49 (35%) gusA sites of integration contained one copy of the gene. Only 11 gusA sites (22%) were linked to the site of integration of the selectable marker. When one site of integration contained several copies of the transgene, a predominance of 3′–3′ inverted re-arrangement repeats was observed. More... »

PAGES

461-473

References to SciGraph publications

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  • 1999-03. Transgene expression in rice engineered through particle bombardment: molecular factors controlling stable expression and transgene silencing in PLANTA
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  • 2001-09-21. Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event in PLANT CELL REPORTS
  • 2000-06. Gene silencing in potato: allelic differences and effect of ploidy in PLANT MOLECULAR BIOLOGY
  • 2000-02. Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns in TRANSGENIC RESEARCH
  • 1999-10-01. Metabolic engineering of Arabidopsis and Brassica for poly(3-hydroxybutyrate- co-3-hydroxyvalerate) copolymer production in NATURE BIOTECHNOLOGY
  • 1993-11. Single-copy T-DNA insertions in Arabidopsis are the predominant form of integration in root-derived transgenics, whereas multiple insertions are found in leaf discs in PLANT MOLECULAR BIOLOGY
  • 1996-09. High-efficiency, microprojectile-mediated cotransformation of sugarcane, using visible or selectable markers in MOLECULAR BREEDING
  • 1998-11. Expression and inheritance of multiple transgenes in rice plants in NATURE BIOTECHNOLOGY
  • 1991-09. Two T-DNA's co-transformed intoBrassica napus by a doubleAgrobacterium tumefaciens infection are mainly integrated at the same locus in THEORETICAL AND APPLIED GENETICS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1024267906219

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    http://dx.doi.org/10.1023/a:1024267906219

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/12885167


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