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Forward genetic approaches in Chlamydia trachomatis.

Publication ,  Journal Article
Nguyen, BD; Valdivia, RH
Published in: J Vis Exp
October 23, 2013

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.

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Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

October 23, 2013

Issue

80

Start / End Page

e50636

Location

United States

Related Subject Headings

  • Sequence Analysis, DNA
  • Mutagenesis
  • Genomics
  • Genome, Bacterial
  • DNA, Bacterial
  • Chlamydia trachomatis
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology
 

Citation

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Nguyen, B. D., & Valdivia, R. H. (2013). Forward genetic approaches in Chlamydia trachomatis. J Vis Exp, (80), e50636. https://doi.org/10.3791/50636
Nguyen, Bidong D., and Raphael H. Valdivia. “Forward genetic approaches in Chlamydia trachomatis.J Vis Exp, no. 80 (October 23, 2013): e50636. https://doi.org/10.3791/50636.
Nguyen BD, Valdivia RH. Forward genetic approaches in Chlamydia trachomatis. J Vis Exp. 2013 Oct 23;(80):e50636.
Nguyen, Bidong D., and Raphael H. Valdivia. “Forward genetic approaches in Chlamydia trachomatis.J Vis Exp, no. 80, Oct. 2013, p. e50636. Pubmed, doi:10.3791/50636.
Nguyen BD, Valdivia RH. Forward genetic approaches in Chlamydia trachomatis. J Vis Exp. 2013 Oct 23;(80):e50636.

Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

October 23, 2013

Issue

80

Start / End Page

e50636

Location

United States

Related Subject Headings

  • Sequence Analysis, DNA
  • Mutagenesis
  • Genomics
  • Genome, Bacterial
  • DNA, Bacterial
  • Chlamydia trachomatis
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology