Genetic Manipulation of Chlamydia trachomatis: Chromosomal Deletions

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

5 Scopus citations

Abstract

Progress in understanding molecular mechanisms contributing to chlamydial pathogenesis has been greatly facilitated by recent advances in genetic manipulation of C. trachomatis. Valuable approaches such as random, chemically induced mutagenesis or targeted, insertion-based gene disruption have led to significant discoveries. We describe herein a technique for generating definitive null strains via complete deletion of chromosomal genes in C. trachomatis. Fluorescence-reported allelic exchange mutagenesis (FRAEM), using the suicide vector pSUmC, enables targeted deletion of desired chromosomal DNA. The protocol provided here describes steps required to produce transformation competent chlamydiae, generate a specific allelic exchange plasmid construct, carry out mutagenesis, and isolate clonal populations of resulting mutant strains.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages151-164
Number of pages14
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume2042
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2019.

Keywords

  • Allelic exchange
  • FRAEM
  • Mutagenesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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