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Updated: Mar 10, 2026

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

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

Published on: November 23, 2012

Markerless Genome Editing in Competent Streptococci.

Roger Junges¹, Rabia Khan¹, Yanina Tovpeko²

¹Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.

Methods in Molecular Biology (Clifton, N.J.)

|December 8, 2016

Summary

This study introduces a markerless genome editing protocol for Streptococcus bacteria using natural genetic transformation. This method enhances precision and avoids unwanted gene accumulation, improving genetic engineering efficiency.

Keywords:

CSP Competence Markerless mutagenesis Natural transformation Pheromone Streptococcus Streptococcus mutans Streptococcus pneumoniae XIP

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Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning

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Published on: October 23, 2017

Area of Science:

Microbiology
Molecular Biology
Bacterial Genetics

Background:

Classical mutagenesis often relies on selective markers, which can interfere with gene expression and lead to unwanted genetic elements.
These limitations hinder precise genetic manipulation in bacteria like Streptococcus mutans and Streptococcus pneumoniae.

Purpose of the Study:

To develop a markerless genome editing protocol for Streptococcus species.
To improve the efficiency and precision of genetic modification through natural transformation.

Main Methods:

Utilizing natural transformation in Streptococcus mutans and Streptococcus pneumoniae.
Inducing competence using specific peptides: XIP for S. mutans and CSP for S. pneumoniae in chemically defined media (CDM).
Employing donor amplicons with extensive flanking homology for precise allelic integration.

Main Results:

Achieved high yields of transformants in both bacterial species.
Demonstrated efficient and precise integration of new alleles without using selective markers.
Successfully combined peptide-induced competence with homologous recombination for markerless editing.

Conclusions:

The presented protocol offers an efficient and precise method for markerless genome editing in Streptococcus.
This approach overcomes limitations of traditional selective markers, enabling cleaner genetic modifications.
Facilitates advanced genetic engineering and research in medically relevant Streptococcus species.