Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis

S. pyogenes secrete a large array of molecules that might contribute to resistance against antimicrobial peptides (AMPs). Some of these are anchored in the cell-wall by enzymes called sortases and others are released from the cell after secretion. One of the released proteins that have previously be...

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Main Authors:	Shahdan, Intan Azura, Smith, Wendy, Kehoe, Mike
Format:	Conference or Workshop Item
Language:	English
Published:	2010
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/15867/ http://irep.iium.edu.my/15867/ http://irep.iium.edu.my/15867/1/construction_of_streptococcus.pdf

id	iium-15867
recordtype	eprints
spelling	iium-158672013-08-22T02:10:51Z http://irep.iium.edu.my/15867/ Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis Shahdan, Intan Azura Smith, Wendy Kehoe, Mike Q Science (General) S. pyogenes secrete a large array of molecules that might contribute to resistance against antimicrobial peptides (AMPs). Some of these are anchored in the cell-wall by enzymes called sortases and others are released from the cell after secretion. One of the released proteins that have previously been reported to inhibit bactericidal effect of the AMPs are streptococcal inhibitor of complement (SIC). SIC has been recognised as a substantial virulence factor in the M1 GAS strain AP1, because Δsic mutant failed to colonise mouse throat (Lukomski et al., 2000). In fact, the colonisation of the SIC-negative strain was significantly impaired during the first four days of post-inoculation, showing that SIC is a crucial virulence factor during the early stages of infection by this strain (Lukomski et al., 2000). Hoe and colleagues then showed that GAS Δsic mutant was easily internalised and killed more effectively by human epithelial cells than the wild-type strain (Hoe et al., 2002). Previous studies used SIC-defective mutants which were constructed by insertion of transposon. The disadvantage of such mutagenesis is that the GAS mutants may have retained an intact copy of the sic gene, which could express some SIC protein. Therefore, the specific objective of the project was to construct a sic deletion mutant of GAS strain SF370 by allele-replacement mutagenesis. The constructed mutant would have a genetically clearly-defined deletion mutant, that completely removed sic gene from the S. pyogenes strain SF370 chromosome, without leaving any foreign sequences (such as an inserted plasmid) behind. 2010 Conference or Workshop Item NonPeerReviewed application/pdf en http://irep.iium.edu.my/15867/1/construction_of_streptococcus.pdf Shahdan, Intan Azura and Smith, Wendy and Kehoe, Mike (2010) Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis. In: IIUM Research, Innovation & Invention Exhibition (IRIIE 2010), 26 - 27 January 2010, Kuala Lumpur. http://www.iium.edu.my/irie/10/info/Programme_Book%20Part_2.pdf
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General)
spellingShingle	Q Science (General) Shahdan, Intan Azura Smith, Wendy Kehoe, Mike Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
description	S. pyogenes secrete a large array of molecules that might contribute to resistance against antimicrobial peptides (AMPs). Some of these are anchored in the cell-wall by enzymes called sortases and others are released from the cell after secretion. One of the released proteins that have previously been reported to inhibit bactericidal effect of the AMPs are streptococcal inhibitor of complement (SIC). SIC has been recognised as a substantial virulence factor in the M1 GAS strain AP1, because Δsic mutant failed to colonise mouse throat (Lukomski et al., 2000). In fact, the colonisation of the SIC-negative strain was significantly impaired during the first four days of post-inoculation, showing that SIC is a crucial virulence factor during the early stages of infection by this strain (Lukomski et al., 2000). Hoe and colleagues then showed that GAS Δsic mutant was easily internalised and killed more effectively by human epithelial cells than the wild-type strain (Hoe et al., 2002). Previous studies used SIC-defective mutants which were constructed by insertion of transposon. The disadvantage of such mutagenesis is that the GAS mutants may have retained an intact copy of the sic gene, which could express some SIC protein. Therefore, the specific objective of the project was to construct a sic deletion mutant of GAS strain SF370 by allele-replacement mutagenesis. The constructed mutant would have a genetically clearly-defined deletion mutant, that completely removed sic gene from the S. pyogenes strain SF370 chromosome, without leaving any foreign sequences (such as an inserted plasmid) behind.
format	Conference or Workshop Item
author	Shahdan, Intan Azura Smith, Wendy Kehoe, Mike
author_facet	Shahdan, Intan Azura Smith, Wendy Kehoe, Mike
author_sort	Shahdan, Intan Azura
title	Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
title_short	Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
title_full	Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
title_fullStr	Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
title_full_unstemmed	Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis
title_sort	construction of streptococcus pyogenes mutants by allele replacement mutagenesis
publishDate	2010
url	http://irep.iium.edu.my/15867/ http://irep.iium.edu.my/15867/ http://irep.iium.edu.my/15867/1/construction_of_streptococcus.pdf
first_indexed	2023-09-18T20:24:46Z
last_indexed	2023-09-18T20:24:46Z
_version_	1777408343002841088

Construction of Streptococcus pyogenes mutants by allele replacement mutagenesis

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