Virulence determinants in the obligate intracellular pathogen Chlamydia trachomatis revealed by forward genetic approaches.
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.
Department of Molecular Genetics and Microbiology, Center for Microbial Pathogenesis, Duke University Medical Center, Durham, NC 27710, USA.
Bidong D
BD
Nguyen
Raphael H
RH
Valdivia
D000818:Animals; D001483:Base Sequence; D002692:Chlamydia trachomatis / Q000235:genetics* / Q000472:pathogenicity*; D002522:Chlorocebus aethiops; D015723:Gene Library; D005821:Genetic Techniques; D023281:Genomics / Q000379:methods; D005838:Genotype; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D014709:Vero Cells; D037521:Virulence Factors / Q000235:genetics*
D037521:Virulence Factors