Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement

Timothy C Barnett; Jessica N Daw; Mark J Walker; Stephan Brouwer

doi:10.1007/978-1-0716-0467-0_5

Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement

Timothy C Barnett, Jessica N Daw, Mark J Walker, Stephan Brouwer

Research output: Chapter in Book/Conference paper › Chapter

10 Citations (Web of Science)

Abstract

Genetic manipulation of Streptococcus pyogenes (Group A Streptococcus, GAS) has historically been a challenging process, with considerable variation in efficiency between different strains. Here, we outline an optimized, rapid method for creating markerless isogenic mutations that combines Gibson assembly cloning with a new temperature-sensitive plasmid, pLZts. This method is highly efficient and reduces the time needed to create GAS mutants to ~2-3 weeks, with the ability to prepare multiple mutants simultaneously.

Original language	English
Title of host publication	Group A Streptococcus
Subtitle of host publication	Methods and Protocols
Editors	Thomas Proft, Jacelyn M. S. Loh
Place of Publication	USA
Publisher	Humana Press Inc.
Chapter	5
Pages	59-69
Number of pages	11
Volume	2136
ISBN (Electronic)	9781071604670
ISBN (Print)	9781071604663
DOIs	https://doi.org/10.1007/978-1-0716-0467-0_5
Publication status	Published - 2020

Publication series

Name	Methods in Molecular Biology
Volume	2136
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Access to Document

10.1007/978-1-0716-0467-0_5

Cite this

@inbook{1082f84e89644c9e9f8af8e139469ace,

title = "Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement",

abstract = "Genetic manipulation of Streptococcus pyogenes (Group A Streptococcus, GAS) has historically been a challenging process, with considerable variation in efficiency between different strains. Here, we outline an optimized, rapid method for creating markerless isogenic mutations that combines Gibson assembly cloning with a new temperature-sensitive plasmid, pLZts. This method is highly efficient and reduces the time needed to create GAS mutants to \textasciitilde{}2-3 weeks, with the ability to prepare multiple mutants simultaneously.",

author = "Barnett, \{Timothy C\} and Daw, \{Jessica N\} and Walker, \{Mark J\} and Stephan Brouwer",

year = "2020",

doi = "10.1007/978-1-0716-0467-0\_5",

language = "English",

isbn = "9781071604663",

volume = "2136",

series = "Methods in Molecular Biology",

publisher = "Humana Press Inc.",

pages = "59--69",

editor = "Proft, \{Thomas \} and Loh, \{Jacelyn M. S. \}",

booktitle = "Group A Streptococcus",

address = "United States",

}

Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement. / Barnett, Timothy C; Daw, Jessica N; Walker, Mark J et al.
Group A Streptococcus: Methods and Protocols. ed. / Thomas Proft; Jacelyn M. S. Loh. Vol. 2136 USA: Humana Press Inc., 2020. p. 59-69 (Methods in Molecular Biology; Vol. 2136).

Research output: Chapter in Book/Conference paper › Chapter

TY - CHAP

T1 - Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement

AU - Barnett, Timothy C

AU - Daw, Jessica N

AU - Walker, Mark J

AU - Brouwer, Stephan

PY - 2020

Y1 - 2020

N2 - Genetic manipulation of Streptococcus pyogenes (Group A Streptococcus, GAS) has historically been a challenging process, with considerable variation in efficiency between different strains. Here, we outline an optimized, rapid method for creating markerless isogenic mutations that combines Gibson assembly cloning with a new temperature-sensitive plasmid, pLZts. This method is highly efficient and reduces the time needed to create GAS mutants to ~2-3 weeks, with the ability to prepare multiple mutants simultaneously.

AB - Genetic manipulation of Streptococcus pyogenes (Group A Streptococcus, GAS) has historically been a challenging process, with considerable variation in efficiency between different strains. Here, we outline an optimized, rapid method for creating markerless isogenic mutations that combines Gibson assembly cloning with a new temperature-sensitive plasmid, pLZts. This method is highly efficient and reduces the time needed to create GAS mutants to ~2-3 weeks, with the ability to prepare multiple mutants simultaneously.

UR - https://www.scopus.com/pages/publications/85084962869

U2 - 10.1007/978-1-0716-0467-0_5

DO - 10.1007/978-1-0716-0467-0_5

M3 - Chapter

C2 - 32430813

SN - 9781071604663

VL - 2136

T3 - Methods in Molecular Biology

SP - 59

EP - 69

BT - Group A Streptococcus

A2 - Proft, Thomas

A2 - Loh, Jacelyn M. S.

PB - Humana Press Inc.

CY - USA

ER -

Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement

Abstract

Publication series

Access to Document

Other files and links

Fingerprint

Improving Health Outcomes in the Tropical North - A Multidisciplinary Collaboration (HOT NORTH)

Cite this

Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement

Abstract

Publication series

Access to Document

Other files and links

Fingerprint

Projects

Improving Health Outcomes in the Tropical North - A Multidisciplinary Collaboration (HOT NORTH)

Cite this