Effective production of marker-free transgenic strawberry plants using inducible site-specific recombination and a bifunctional selectable marker gene

J.G. Schaart, F.A. Krens, K.T.B. Pelgrom, O. Mendes, G.J.A. Rouwendal

Research output: Contribution to journalArticleAcademicpeer-review

115 Citations (Scopus)

Abstract

Public concerns about the issue of the environmental safety of genetically modified plants have led to a demand for technologies allowing the production of transgenic plants without selectable (antibiotic resistance) markers. We describe the development of an effective transformation system for generating such marker-free transgenic plants, without the need for repeated transformation or sexual crossing. This system combines an inducible site-specific recombinase for the precise elimination of undesired, introduced DNA sequences with a bifunctional selectable marker gene used for the initial positive selection of transgenic tissue and subsequent negative selection for fully marker-free plants. The described system can be generally applied to existing transformation protocols, and was tested in strawberry using a model vector in which site-specific recombination leads to a functional combination of a cauliflower mosaic virus 35S promoter and a GUS encoding sequence, thereby enabling the histochemical monitoring of recombination events. Fully marker-free transgenic strawberry plants were obtained following two different selection/regeneration strategies.
Original languageEnglish
Pages (from-to)233-240
JournalPlant Biotechnology Journal
Volume2
Issue number3
DOIs
Publication statusPublished - 2004

Fingerprint

site-specific recombination
Fragaria
Genetically Modified Plants
Genetic Recombination
strawberries
transgenic plants
genetically modified organisms
genetic markers
Cauliflower mosaic virus
antibiotic resistance
Genes
production technology
promoter regions
Caulimovirus
nucleotide sequences
monitoring
Microbial Drug Resistance
Regeneration
Technology
Safety

Keywords

  • cytosine deaminase
  • glucocorticoid receptor
  • transformation vector
  • negative selection
  • bacterial gene
  • expression
  • agrobacterium
  • regeneration
  • system
  • frequency

Cite this

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title = "Effective production of marker-free transgenic strawberry plants using inducible site-specific recombination and a bifunctional selectable marker gene",
abstract = "Public concerns about the issue of the environmental safety of genetically modified plants have led to a demand for technologies allowing the production of transgenic plants without selectable (antibiotic resistance) markers. We describe the development of an effective transformation system for generating such marker-free transgenic plants, without the need for repeated transformation or sexual crossing. This system combines an inducible site-specific recombinase for the precise elimination of undesired, introduced DNA sequences with a bifunctional selectable marker gene used for the initial positive selection of transgenic tissue and subsequent negative selection for fully marker-free plants. The described system can be generally applied to existing transformation protocols, and was tested in strawberry using a model vector in which site-specific recombination leads to a functional combination of a cauliflower mosaic virus 35S promoter and a GUS encoding sequence, thereby enabling the histochemical monitoring of recombination events. Fully marker-free transgenic strawberry plants were obtained following two different selection/regeneration strategies.",
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Effective production of marker-free transgenic strawberry plants using inducible site-specific recombination and a bifunctional selectable marker gene. / Schaart, J.G.; Krens, F.A.; Pelgrom, K.T.B.; Mendes, O.; Rouwendal, G.J.A.

In: Plant Biotechnology Journal, Vol. 2, No. 3, 2004, p. 233-240.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Schaart, J.G.

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AU - Pelgrom, K.T.B.

AU - Mendes, O.

AU - Rouwendal, G.J.A.

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