Virulence Determinants in the Obligate Intracellular Pathogen Chlamydia Trachomatis Revealed by Forward Genetic Approaches

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2012 Jan 11

PMID 22232666

Citations 88

Authors

Bidong D Nguyen

Raphael H Valdivia

Affiliations

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Abstract

Chlamydia trachomatis, a pathogen responsible for diseases of significant clinical and public health importance, remains poorly characterized because of its intractability to routine molecular genetic manipulation. We have developed a combinatorial approach to rapidly generate a comprehensive library of genetically defined mutants. Chemical mutagenesis, coupled with whole-genome sequencing (WGS) and a system for DNA exchange within infected cells, was used to generate Chlamydia mutants with distinct phenotypes, map the underlying genetic lesions, and generate isogenic strains. As a result, we identified mutants with altered glycogen metabolism, including an attenuated strain defective for type II secretion. The coupling of chemically induced gene variation and WGS to establish genotype-phenotype associations should be broadly applicable to the large list of medically and environmentally important microorganisms currently intractable to genetic analysis.

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