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Virulence determinants in the obligate intracellular pathogen Chlamydia trachomatis revealed by forward genetic approaches.


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.

SUBMITTER: Nguyen BD 

PROVIDER: S-EPMC3268281 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature

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Virulence determinants in the obligate intracellular pathogen Chlamydia trachomatis revealed by forward genetic approaches.

Nguyen Bidong D BD   Valdivia Raphael H RH  

Proceedings of the National Academy of Sciences of the United States of America 20120109 4


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 dist  ...[more]

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