Transposon mutagenesis in clostridium difficile

Haitham A. Hussain; Adam Roberts; Rachael Whalan; Peter Mullany

doi:10.1007/978-1-60327-365-7_13

Transposon mutagenesis in clostridium difficile

Haitham A. Hussain, Adam Roberts, Rachael Whalan, Peter Mullany

University College London

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

6 Citations (Scopus)

Abstract

Genetic manipulation of Clostridium difficile is notoriously difficult, currently there is only one reliable method for generating random mutations in the organism and that is to use the conjugative transposon Tn916. Tn916 enters the genome of most strains of C. difficile with no obvious target site preference. In order to use the genome strain C. difficile 630 for transposon mutagenesis a erythromycin-sensitive derivative C. difficile 630Δerm was constructed and the Tn916 derivative, Tn916ΔE, was shown to enter the genome at multiple sites enabling the construction of a Tn916 insertion library.

Original language	English
Title of host publication	Methods in Molecular Biology
Editors	Peter Mullany, Adam Roberts
Pages	203-211
Number of pages	9
DOIs	https://doi.org/10.1007/978-1-60327-365-7_13
Publication status	Published - 1 Jan 2010
Externally published	Yes

Keywords

630Δerm
Clostridium difficile
mutant library
Tn916
transposon mutagenesis

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10.1007/978-1-60327-365-7_13

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abstract = "Genetic manipulation of Clostridium difficile is notoriously difficult, currently there is only one reliable method for generating random mutations in the organism and that is to use the conjugative transposon Tn916. Tn916 enters the genome of most strains of C. difficile with no obvious target site preference. In order to use the genome strain C. difficile 630 for transposon mutagenesis a erythromycin-sensitive derivative C. difficile 630Δerm was constructed and the Tn916 derivative, Tn916ΔE, was shown to enter the genome at multiple sites enabling the construction of a Tn916 insertion library.",

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AB - Genetic manipulation of Clostridium difficile is notoriously difficult, currently there is only one reliable method for generating random mutations in the organism and that is to use the conjugative transposon Tn916. Tn916 enters the genome of most strains of C. difficile with no obvious target site preference. In order to use the genome strain C. difficile 630 for transposon mutagenesis a erythromycin-sensitive derivative C. difficile 630Δerm was constructed and the Tn916 derivative, Tn916ΔE, was shown to enter the genome at multiple sites enabling the construction of a Tn916 insertion library.

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BT - Methods in Molecular Biology

A2 - Mullany, Peter

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ER -

Transposon mutagenesis in clostridium difficile

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Keywords

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