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The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker
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Biologia
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JoVE Journal Biologia
The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

DOI:
10.3791/2331-v

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12:03 min

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January 11, 2011

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, , ,
1Department of Radiation Oncology,Beth Israel Deaconess Medical Center, Harvard Medical School, 2Department of Medicine, Division of Geriatric Medicine and Pittsburgh Institute for Neurodegenerative Diseases,University of Pittsburgh

Capítulos

  • 00:05Título
  • 01:47Transfer Fosmid to SW016 Bacteria
  • 04:49Insertion of the galK Gene by Recombineering
  • 08:48Replacement of galK with Tag Sequences by Recombineering
  • 10:48Representative Results
  • 11:34Conclusion

Summary

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The ability to produce transgenes for Caenorhabditis elegans using genomic DNA carried by fosmids is particularly attractive as all of the native regulatory elements are retained. Described is a simple and robust procedure for the production of transgenes via recombineering with the galK selectable marker.

Tags

C. ElegansTransgenesRecombineeringGalK Selectable MarkerGFP Fusion ProteinsGene RegulationExpression PatternTandem Affinity Purification (TAP)PromoterReporter GeneCDNA3′ Untranslated RegionAlternative PromotersWorm Genomic DNAFosmid DNA ClonesE. Coli Based Recombineering ProcedureGFP InsertionTAP-tag InsertionPositive SelectionNegative Selection

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