Improved vectors for selection of transgenic caenorhabditis elegans

Annabel A. Ferguson, Liquan Cai, Luv Kashyap, Alfred L. Fisher

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

Abstract

The generation of transgenic animals is an essential part of research in Caenorhabditis elegans. One technique for the generation of these animals is biolistic bombardment involving the use of DNA-coated microparticles. To facilitate the identification of transgenic animals within a background of non-transformed animals, the unc-119 gene is often used as a visible marker as the unc-119 mutants are small and move poorly and the larger size and smoother movement of rescued animals make them clearly visible. While transgenic animals can be identified from co-bombardment with a transgene of interest and a separate unc-119 rescue plasmid, placing the unc-119 in cis on the transgene increases confidence that the resulting transgenic animals contain and express both the marker and the transgene. However, placing the unc-119 marker on the backbone of a plasmid or larger DNA construct, such as a fosmid or BAC, can be technically difficult using standard molecular biology techniques. Here we describe methods to circumvent these limitations and use either homologous recombination or Cre-LoxP mediated recombination in Escherichia coli to insert the unc-119 marker on to a variety of vector backbones.

Original languageEnglish (US)
Title of host publicationBiolistic DNA Delivery
Subtitle of host publicationMethods and Protocols
EditorsStephan Sudowe, Angelika Reske-Kunz
Pages87-102
Number of pages16
DOIs
StatePublished - Jan 1 2013

Publication series

NameMethods in Molecular Biology
Volume940
ISSN (Print)1064-3745

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Keywords

  • Biotechnology
  • C. elegans
  • Cre recombinase
  • Microparticle bombardment
  • Plasmid
  • Recombination
  • Transgenic animals
  • unc-119

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Ferguson, A. A., Cai, L., Kashyap, L., & Fisher, A. L. (2013). Improved vectors for selection of transgenic caenorhabditis elegans. In S. Sudowe, & A. Reske-Kunz (Eds.), Biolistic DNA Delivery: Methods and Protocols (pp. 87-102). (Methods in Molecular Biology; Vol. 940). https://doi.org/10.1007/978-1-62703-110-3_8