title: Forward genetic approaches in Chlamydia trachomatis.
abstract: Chlamydia trachomatis, the etiological agent of sexually transmitted diseases and ocular infections, remains poorly characterized due to its intractability to experimental transformation with recombinant DNA. We developed an approach to perform genetic analysis in C. trachomatis despite the lack of molecular genetic tools. Our method involves: i.) chemical mutagenesis to rapidly generate comprehensive libraries of genetically-defined mutants with distinct phenotypes; ii.) whole-genome sequencing (WGS) to map the underlying genetic lesions and to find associations between mutated gene(s) and a common phenotype; iii.) generation of recombinant strains through co-infection of mammalian cells with mutant and wild type bacteria. Accordingly, we were able to establish causal relationships between genotypes and phenotypes. The coupling of chemically-induced gene variation and WGS to establish correlative genotype-phenotype associations should be broadly applicable to the large list of medically and environmentally important microorganisms currently intractable to genetic analysis.
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Microbial Pathogenesis, Duke University Medical Center.
authors.forename: Bidong D
authors.initials: BD
authors.lastname: Nguyen
authors.forename: Raphael H
authors.initials: RH
authors.lastname: Valdivia
mesh_terms: D002692:Chlamydia trachomatis / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods*; D016296:Mutagenesis; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004269:DNA, Bacterial