The cre/lox system: A practical tool to efficiently eliminate selectable markers in fungal endophytes

Simona Florea, Caroline MacHado, Kalina Andreeva, Christopher L. Schardl

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

Interspecific hybridization is a common evolutionary process for the many epichloid fungi that consequently possess multiple gene copies accumulated from their parental strains. Serial gene manipulations in such strains are impeded by the limited availability of selectable resistance marker genes. Therefore, we developed a method for marker elimination suitable for a range of filamentous fungi that allows the reuse of the same marker for successive manipulations, and can also generate gene knockout mutants free of any foreign genes. For epichloae, the complete elimination of the marker gene from the genome would mitigate public concerns and regulatory hurdles to the use of such fungal strains in field experiments.

Original languageEnglish
Title of host publicationRecombinant Gene Expression
Subtitle of host publicationReviews and Protocols, Third Edition
EditorsArgelia Lorence
Pages371-379
Number of pages9
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume824
ISSN (Print)1064-3745

Keywords

  • Cre/lox
  • Endophyte
  • Epichloë
  • Marker excision
  • Transient expression
  • Transient transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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