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Updated: Jul 3, 2026

Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning
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Published on: October 23, 2017

Shuttle expression plasmids for genetic studies in Streptococcus mutans.

Indranil Biswas1, Jyoti K Jha1, Nicholas Fromm1

  • 1Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA.

Microbiology (Reading, England)
|August 1, 2008
PubMed
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Researchers developed new shuttle plasmids for gene overexpression in streptococci. These plasmids utilize broad-host-range replicons and constitutive promoters for versatile gene expression and recombinant protein production in bacteria.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Streptococci, including Streptococcus mutans, are important targets for genetic manipulation.
  • Efficient gene expression systems are crucial for studying bacterial functions and developing biotechnological applications.

Purpose of the Study:

  • To create a versatile set of shuttle plasmids for the overexpression of genes in streptococci and other Gram-positive bacteria.
  • To provide tools for complementation assays and recombinant protein expression.

Main Methods:

  • Construction of shuttle plasmids based on rolling-circle (pSH71) and theta (pAMbeta1) replicons.
  • Incorporation of four constitutive promoters (P(ami), P(spac), P(23), P(veg)) for varied gene expression levels.
  • Generation of plasmids for N-terminal 6xHis or C-terminal Strep-tag fusion protein expression.

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Published on: November 23, 2012

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Last Updated: Jul 3, 2026

Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning
12:06

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

Published on: October 23, 2017

Purification of a High Molecular Mass Protein in Streptococcus mutans
09:51

Purification of a High Molecular Mass Protein in Streptococcus mutans

Published on: September 14, 2019

Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR
14:07

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

Published on: November 23, 2012

Main Results:

  • Plasmids demonstrated utility in a gene complementation assay.
  • High-level recombinant protein expression was achieved in Streptococcus mutans and Streptococcus pyogenes using the developed plasmids.
  • Theta-replicons were chosen to mitigate stability issues associated with rolling-circle replicons during large gene cloning.

Conclusions:

  • The developed shuttle plasmids offer a flexible platform for gene expression studies in a wide range of Gram-positive bacteria.
  • These tools facilitate both gene complementation and the production of tagged recombinant proteins.
  • The broad-host-range nature and functional promoters enable diverse applications in bacterial genetics and biotechnology.