Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Global genome analysis identifies glycolipids and lipoteichoic acid alanylation as contributors to group B streptococcal diabetic wound infection.

Cell reports·2025
Same author

Global genome analysis identifies glycolipids and lipoteichoic acid alanylation as contributors to Group B streptococcal diabetic wound infection.

bioRxiv : the preprint server for biology·2025
Same author

Group A Streptococcal asparagine metabolism regulates bacterial virulence.

EMBO reports·2025
Same author

Identification of glyoxalase A in group B <i>Streptococcus</i> and its contribution to methylglyoxal tolerance and virulence.

Infection and immunity·2025
Same author

The PdxR-PdxKU locus involved in vitamin B<sub>6</sub> salvage is important for group A streptococcal resistance to neutrophil killing and survival in human blood.

Microbiology spectrum·2024
Same author

da_Tracker: Automated workflow for high throughput single cell and single phagosome tracking in infected cells.

Biology open·2024

Related Experiment Video

Updated: Dec 21, 2025

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

16.9K

Protocols for Tn-seq Analyses in the Group A Streptococcus.

Yoann Le Breton1, Ashton T Belew2,3, Kevin S McIver2

  • 1Wound Infections Department, Bacterial Diseases Branch, Walter Reed Army Institute for Research, Silver Spring, MD, USA. yoann.s.lebreton.ctr@mail.mil.

Methods in Molecular Biology (Clifton, N.J.)
|May 21, 2020
PubMed
Summary
This summary is machine-generated.

Transposon-sequencing (Tn-seq) provides a powerful method for studying bacterial genetics. This chapter details Tn-seq protocols for Streptococcus pyogenes, enabling genome-wide analysis of mutant libraries.

Keywords:
Isolate 5448Krmit transposonMarinerTn-seq

More Related Videos

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
08:34

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

Published on: February 23, 2021

7.3K
Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
08:19

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Published on: July 7, 2020

11.2K

Related Experiment Videos

Last Updated: Dec 21, 2025

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

16.9K
MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
08:34

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

Published on: February 23, 2021

7.3K
Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
08:19

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Published on: July 7, 2020

11.2K

Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Transposon-sequencing (Tn-seq) is a powerful tool for bacterial genetics, enabling large-scale analysis of mutant libraries.
  • Understanding genotype-phenotype associations and bacterial physiology requires high-resolution, genome-wide methods.
  • Streptococcus pyogenes (Group A Streptococcus or GAS) is a significant human pathogen, necessitating advanced genetic analysis tools.

Purpose of the Study:

  • To present detailed, step-by-step protocols for performing Tn-seq analyses in Streptococcus pyogenes.
  • To enable the generation of complex Krmit transposon mutant libraries in GAS.
  • To facilitate the sequencing and analysis of transposon-genome junctions for Tn-seq studies.

Main Methods:

  • Generation of highly complex Krmit transposon mutant libraries in GAS.
  • En masse production of Krmit insertion tags for high-throughput sequencing.
  • Illumina sequencing of transposon-genome junctions for Tn-seq data acquisition.
  • Application of established protocols to various GAS serotypes and other pathogenic Streptococci.

Main Results:

  • Successful generation of complex Krmit mutant libraries in S. pyogenes.
  • Detailed methodology for preparing transposon insertion libraries for sequencing.
  • Demonstrated applicability of the protocols across multiple GAS serotypes.
  • Established a robust Tn-seq pipeline for genetic analysis in pathogenic Streptococci.

Conclusions:

  • The presented protocols provide a standardized approach for Tn-seq in Streptococcus pyogenes.
  • This methodology facilitates genome-scale genetic studies of GAS and related pathogens.
  • Tn-seq analysis of GAS mutant libraries enhances understanding of bacterial physiology and pathogenesis.
  • The Krmit transposon system offers a versatile tool for forward genetics in Streptococci.