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Related Experiment Videos

Site-specific recombination in Zea mays.

S Kerbach1, H Lörz, D Becker

  • 1Biocenter Klein Flottbek, Section Developmental Biology and Biotechnology, University of Hamburg, Ohnhorststrasse 18, 22609, Hamburg, Germany.

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik
|October 4, 2005
PubMed
Summary

Site-specific recombination systems efficiently eliminate marker genes in genetically modified maize. The Cre/lox system precisely removed the pat gene, while Flp/FRT showed limited effectiveness, indicating Cre/lox is superior for commercial applications.

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Area of Science:

  • Plant biotechnology
  • Molecular genetics
  • Genetically modified organisms

Background:

  • Marker gene elimination is crucial for commercializing genetically modified (GM) plants.
  • Site-specific recombination (SSR) systems offer precise gene excision.
  • Cre/lox and Flp/FRT are common SSR systems with varying efficiencies.

Purpose of the Study:

  • To compare the efficacy of Cre/lox and Flp/FRT systems for marker gene elimination in maize.
  • To assess the potential for directed integration of genetic elements using SSR systems.

Main Methods:

  • Maize plants were engineered to contain the pat selection marker gene flanked by lox or FRT sites.
  • Sexual crossing was employed to introduce constitutively expressed Cre or Flp recombinases.

Related Experiment Videos

  • Biolistic bombardment was used to test site-specific integration with a lox.uidA construct.

    • Main Results:

    • Cre/lox mediated precise and complete excision of the lox-flanked marker gene in F1 progeny.
    • Flp-mediated recombination of FRT sites occurred rarely, indicating lower efficiency.
    • Biolistic bombardment demonstrated directed integration of the lox.uidA sequence into maize embryos.

    Conclusions:

    • The Cre/lox system is highly effective for marker gene elimination in maize, suitable for commercial applications.
    • The Flp/FRT system demonstrated limited utility for marker gene removal in this context.
    • SSR systems, particularly Cre/lox, show promise for both gene elimination and targeted gene insertion in maize.