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Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43
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Immunology and Infection
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JoVE Journal Immunology and Infection
Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43

Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43

DOI:
10.3791/61594-v

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06:06 min

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September 11, 2020

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, , , ,
1Faculty of Infections and Tropical Diseases, Department of Infection Biology,London School of Hygiene and Tropical Medicine, 2Division of Medicine,University College London, 3Division of Infection and Immunity,University College London UCL

Chapters

  • 00:05Introduction
  • 00:35Generation of Plasmid pSD1 and Chemical Transformation of E. coli Dh5α
  • 02:03Plasmid DNA Extraction, Restriction Digestion of pSD1 and Spectinomycin Gene, and Assembly of pSD2
  • 03:03Transformation of S. pneumoniae Strain 519/43
  • 04:46Results: Confirmation of Mutagenesis
  • 05:43Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a S. pneumoniae serotype 1 strain 519/43 that can be genetically modified by using its ability to naturally acquire DNA and a suicide-plasmid. As proof of principle, an isogenic mutant in the pneumolysin (ply) gene was made.

Tags

Keywords: Serotype 1 Streptococcus PneumoniaeGenetic ModificationPlasmid ConstructionPSD1PSD2Spectinomycin CassetteRestriction DigestionLigationE. Coli TransformationS. Pneumoniae Culture

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