Transformation of Plants with Multiple Cassettes Generates Simple Transgene Integration Patterns and High Expression Levels View Full Text


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Article Info

DATE

2005-10

AUTHORS

Pawan K. Agrawal, Ajay Kohli, Richard M. Twyman, Paul Christou

ABSTRACT

We transformed rice (Oryza sativa L.) simultaneously with five minimal cassettes, each containing a promoter, coding region and polyadenylation site but no vector backbone. We found that multi-transgene cotransformation was achieved with high efficiency using multiple cassettes, with all transgenic plants we generated containing at least two transgenes and 16% containing all five. About 75% of the plants had simple transgene integration patterns with a predominance of single-copy insertions. The expression levels for all transgenes, and the overall coexpression frequencies, were much higher than previously reported in whole plasmid transformants. Four of five lines analyzed for transgene expression stability in subsequent generations showed stable and high expression levels over generations. A simple model is proposed, which accounts for differences in the molecular make-up and the expression profile of transgenic plants generated using whole plasmid or minimal cassettes. We conclude that gene transfer using minimal cassettes is an efficient and rapid method for the production of transgenic plants containing and stably expressing several different transgenes. Our results facilitate effective manipulation of multi-gene pathways in plants in a single transformation step. More... »

PAGES

247-260

References to SciGraph publications

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  • 2003-05. Transgene integration, organization and interaction in plants in PLANT MOLECULAR BIOLOGY
  • 1998-07. An Efficient Rice Transformation System Utilizing Mature Seed-derived Explants and a Portable, Inexpensive Particle Bombardment Device in TRANSGENIC RESEARCH
  • 2000-08. Transgenic rice as a system to study the stability of transgene expression: multiple heterologous transgenes show similar behaviour in diverse genetic backgrounds in THEORETICAL AND APPLIED GENETICS
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  • 1993-03. An improved rice transformation system using the biolistic method in PLANT CELL REPORTS
  • 2001-02. Post-transcriptional gene silencing by double-stranded RNA in NATURE REVIEWS GENETICS
  • 2001-07. High-resolution structural analysis of biolistic transgene integration into the genome of wheat in THEORETICAL AND APPLIED GENETICS
  • 2000-02. Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns in TRANSGENIC RESEARCH
  • 1986-12-01. Co–Transformation of Unlinked Foreign Genes into Plants by Direct Gene Transfer in NATURE BIOTECHNOLOGY
  • 1993-03. Use of bar as a selectable marker gene and for the production of herbicide-resistant rice plants from protoplasts in PLANT MOLECULAR BIOLOGY
  • 1995-01. Inheritance of gusA and neo genes in transgenic rice in PLANT MOLECULAR BIOLOGY
  • 1998-11. Expression and inheritance of multiple transgenes in rice plants in NATURE BIOTECHNOLOGY
  • 1990-05. Cotransformation frequencies of foreign genes in soybean cell cultures in THEORETICAL AND APPLIED GENETICS
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    URI

    http://scigraph.springernature.com/pub.10.1007/s11032-005-0239-5

    DOI

    http://dx.doi.org/10.1007/s11032-005-0239-5

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1023530783


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