A Toolkit and Robust Pipeline for the Generation of Fosmid-Based Reporter Genes in C. elegans
2009

A Toolkit for Creating Reporter Genes in C. elegans

publication Evidence: moderate

Author Information

Author(s): Tursun Baris, Cochella Luisa, Carrera Inés, Hobert Oliver

Primary Institution: Howard Hughes Medical Institute, Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, New York, New York, United States of America

Hypothesis

Can a robust pipeline for generating fosmid-based reporter genes improve gene expression studies in C. elegans?

Conclusion

The new recombineering pipeline simplifies and enhances the generation of reporter constructs in C. elegans, allowing for more accurate studies of gene expression.

Supporting Evidence

  • The toolkit allows for the insertion of various fluorescent proteins and affinity tags at specific target sites within fosmid clones.
  • The new pipeline is less complex and works more robustly than previously described methods.
  • Fosmid recombineering provides a way to study gene expression with greater accuracy by including more regulatory information.

Takeaway

This study shows a new way to make special tags that help scientists see how genes work in tiny worms called C. elegans.

Methodology

The study describes a pipeline for recombineering fosmids to create reporter constructs using homologous recombination in bacteria.

Limitations

The efficiency of the recombineering process can vary, and the presence of false positives may complicate the selection of successful recombinants.

Digital Object Identifier (DOI)

10.1371/journal.pone.0004625

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