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Related Experiment Video

Updated: Dec 11, 2025

Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43

Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43

Published on: September 11, 2020

Advancing Genetic Tools in Streptococcus pneumoniae.

Haley Echlin¹, Jason W Rosch¹

¹Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.

|August 23, 2020

Summary

Researchers improved genetic tools for studying Streptococcus pneumoniae. Novel constructs enhance gene deletion efficiency, aiding research into pneumococcal diseases and pathogen manipulation.

Keywords:

Streptococcus pneumoniae allelic replacement markerless cassette

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Last Updated: Dec 11, 2025

Constructing Mutants in Serotype 1 Streptococcus pneumoniae strain 519/43

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Published on: September 11, 2020

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR

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Published on: November 23, 2012

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae

A Fast and Reliable Pipeline for Bacterial Transcriptome Analysis Case study: Serine-dependent Gene Regulation in Streptococcus pneumoniae

Published on: April 25, 2015

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Streptococcus pneumoniae causes numerous diseases, necessitating further study of its pathogenesis.
Genetic manipulation tools exist for S. pneumoniae, but their efficiency can be limiting.

Purpose of the Study:

To evaluate the efficiency of the Sweet Janus cassette for allelic replacement in S. pneumoniae.
To develop novel genetic constructs for improved gene deletion and allelic replacement in S. pneumoniae.

Main Methods:

Utilized the Sweet Janus cassette for capsule locus replacement.
Developed and tested novel cassettes with alternative selection markers, including chlorinated phenylalanine.
Created constructs with single and dual selection markers, some not requiring engineered resistance.

Main Results:

Initial attempts with Sweet Janus showed low allelic replacement efficiency and high false-positive revertants.
Reversion frequency was found to be dependent on the bacterial clone.
Novel dual and triple selection marker cassettes were successfully created and utilized for allelic replacement.

Conclusions:

The efficiency of allelic replacement in S. pneumoniae is influenced by bacterial clone and selection methods.
Novel selection marker cassettes improve gene deletion capabilities in S. pneumoniae.
These new tools expand experimental applications for studying pneumococcal genetics and pathogenesis.