Chlamydia trachomatis is among the most significant bacterial pathogens afflicting humans. Despite this enormous impact on public health, many aspects about C. trachomatis including basic biology, genetics, and pathogenesis are poorly understood. This is reflective of a primary limitation in chlamydial research: the lack of a system for directed genetic manipulation. Two significant achievements were recently reported: a method for introducing DNA into Chlamydia (transformation) and development of a plasmid stable in both E. coli and Chlamydia with an effective selectable antibiotic marker (shuttle vector). This method and tool can enable us, for the first time in the Chlamydia field, the ability to directly introduce desired DNA sequences into Chlamydia and evaluate resulting biological effects, define virulence components, and better understand how disease is caused and ameliorated. Tightly controlled gene expression and targeted gene repression is essential to appropriately analyze the biological effect of a given protein in Chlamydia, especially given the many stages of infection and development. Many molecular components to achieve controlled gene expression and repression have been employed successfully in model bacterial systems;however, none have been tested in Chlamydia.
Three specific aims are proposed;1) Conditional gene expression, 2) Targeted gene repression, and 3) Fluorescent protein validation. Developing these molecular tools in Chlamydia would be pivotal accomplishments in the chlamydial field. Moreover, would allow for the first time the ability to directly demonstrate and define mechanisms for chlamydial pathogenesis.

Public Health Relevance

Chlamydia trachomatis is a medically important bacterium for which basic biology and mechanisms for causing disease are poorly understood. This proposal is designed to develop essential molecular tools enhance our ability to treat and prevent these infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI103711-02
Application #
8605516
Study Section
Special Emphasis Panel (ZRG1-IDM-A (80))
Program Officer
Hiltke, Thomas J
Project Start
2013-01-15
Project End
2014-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
2
Fiscal Year
2014
Total Cost
$164,610
Indirect Cost
$52,110
Name
University of Kansas Lawrence
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
076248616
City
Lawrence
State
KS
Country
United States
Zip Code
66045
Wickstrum, Jason; Sammons, Lindsay R; Restivo, Keasha N et al. (2013) Conditional gene expression in Chlamydia trachomatis using the tet system. PLoS One 8:e76743