Mutagenesis of Chlamydia trachomatis Using TargeTron

Mary M Weber; Robert Faris

doi:10.1007/978-1-4939-9694-0_12

Mutagenesis of Chlamydia trachomatis Using TargeTron

Methods Mol Biol. 2019:2042:165-184. doi: 10.1007/978-1-4939-9694-0_12.

Authors

Mary M Weber¹, Robert Faris²

Affiliations

¹ Department of Microbiology and Immunology, University of Iowa Carver College of Medicine, Iowa City, IA, USA. mary-weber@uiowa.edu.
² Department of Microbiology and Immunology, University of Iowa Carver College of Medicine, Iowa City, IA, USA.

PMID: 31385276
DOI: 10.1007/978-1-4939-9694-0_12

Abstract

Chlamydia trachomatis is an important human pathogen that prior to 2011 was largely intractable to genetic manipulation. Here we describe the application of a group II intron, referred to as TargeTron, for site-specific insertional inactivation of target genetic loci in the obligate, intracellular bacteria C. trachomatis. In this chapter, we outline the methods for intron retargeting, chlamydia transformation, and mutant verification. We also outline a method for complementation of TargeTron mutants. Furthermore, we discuss potential pitfalls and alternative strategies for generating mutants with TargeTron technology.

Keywords: Chlamydia trachomatis; Complementation; Group II intron; Insertional inactivation; TargeTron.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics
Chlamydia Infections / microbiology
Chlamydia trachomatis / genetics*
Gene Targeting / methods*
Humans
Introns
Mutagenesis, Insertional / methods*

Substances

Bacterial Proteins