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Genetic Transformation and Complementation.

D Scott Samuels1, Dan Drecktrah2, Laura S Hall2

  • 1Division of Biological Sciences, University of Montana, Missoula, MT, USA. samuels@mso.umt.edu.

Methods in Molecular Biology (Clifton, N.J.)
|October 17, 2017
PubMed
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Genetic manipulation of Borrelia (Borreliella) burgdorferi is essential for Lyme disease research. This chapter details methods for creating and analyzing bacterial mutants, aiding the study of spirochete pathogenesis.

Area of Science:

  • Microbiology
  • Genetics
  • Pathogenesis

Background:

  • Genetic manipulation of Borrelia (Borreliella) burgdorferi is crucial for understanding Lyme disease.
  • Despite challenges with these recalcitrant bacteria, mutant generation is a routine laboratory practice.

Purpose of the Study:

  • To provide a comprehensive guide to molecular genetic experiments in Borrelia (Borreliella) burgdorferi.
  • To detail methodologies for creating, selecting, and verifying genetic modifications in spirochetes.

Main Methods:

  • Preparation and transformation of competent Borrelia (Borreliella) burgdorferi cells.
  • Selection and isolation techniques for identifying successful transformants.
  • Methods for complementing or genetically restoring mutants to wild-type phenotype.
Keywords:
Antibiotic resistanceBorrelia burgdorferiComplementationElectroporationLyme diseaseMolecular geneticsMutagenesisShuttle vectorSpirocheteTransformation

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Main Results:

  • Established protocols for reproducible genetic manipulation of Borrelia (Borreliella) burgdorferi.
  • Detailed experimental designs for generating and characterizing mutants.
  • Guidance on validating genetic modifications and restoring gene function.

Conclusions:

  • Effective genetic manipulation techniques are vital for advancing Borrelia (Borreliella) burgdorferi microbiology.
  • The methodologies described facilitate the study of Lyme disease pathogenesis through mutant analysis.
  • This chapter serves as a practical resource for researchers working with Borrelia (Borreliella) burgdorferi genetics.