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Methods Mol Biol. 2017;1498:163-177.

Use of Group II Intron Technology for Targeted Mutagenesis in Chlamydia trachomatis.

Author information

1
Department of Microbiology, Southern Illinois University, 1125 Lincoln Drive, Carbondale, IL, 62901, USA.
2
Department of Microbiology, Southern Illinois University, 1125 Lincoln Drive, Carbondale, IL, 62901, USA. dfisher@siu.edu.

Abstract

Dissecting the contribution of genes to virulence in fulfillment of Molecular Koch's postulates is essential for developing prevention and treatment strategies for bacterial pathogens. This chapter will discuss the application of a targeted, intron-based insertional mutagenesis method for creating mutants in the obligate, intracellular bacterial pathogen Chlamydia trachomatis. The methods employed for intron targeting, mutant selection, and mutant verification will be outlined including available selection markers, gene targeting strategies, and potential pitfalls.

KEYWORDS:

Chemical transformation; Chlamydia trachomatis; Gene inactivation; Group II intron; Insertional mutagenesis

PMID:
27709575
DOI:
10.1007/978-1-4939-6472-7_11
[Indexed for MEDLINE]

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