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.
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Acknowledgments
This work was supported by startup funds from the University of Iowa Carver College of Medicine Department of Microbiology and Immunology to M.M.W. We thank Shelby Andersen, Annie Holtz, and Stephanie Peterson for critical review of the manuscript.
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Weber, M.M., Faris, R. (2019). Mutagenesis of Chlamydia trachomatis Using TargeTron. In: Brown, A. (eds) Chlamydia trachomatis. Methods in Molecular Biology, vol 2042. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9694-0_12
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DOI: https://doi.org/10.1007/978-1-4939-9694-0_12
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