Artificial regulation of ospC expression in Borrelia burgdorferi.

Artificial regulation of ospC expression in Borrelia burgdorferi. Research Abstract Details 

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Artificial regulation of ospC expression in Borrelia burgdorferi. Abstract Text:

Michael A Gilbert,Elizabeth A Morton,Sharyl F Bundle,D Scott Samuels,

Outer surface lipoprotein (Osp) C is a virulence factor required for transmission of the Lyme disease agent, Borrelia burgdorferi. We have constructed an inducible promoter system to study the function and regulation of OspC by integrating regulatory elements from the Escherichia coli lac operon into the B. burgdorferi genome. An inducible promoter (flacp) was constructed by inserting a synthetic lac operator sequence between the transcriptional start site and the ribosomal binding site of the B. burgdorferi flgB promoter; flacp was then used to replace the native ospC and rpoS promoters in B. burgdorferi derivatives that constitutively express the E. coli Lac repressor protein (LacI). In vitro, the expression of ospC and rpoS from flacp was dependent on the inducer isopropyl beta-D-thiogalactopyranoside and was unaffected by temperature or pH, conditions commonly used to mimic different aspects of the B. burgdorferi life cycle. Our results suggest that OspC is essential immediately upon injection into a mouse and OspC expression must be maintained during the early stages of infection. In addition, the mouse infectivity experiment indicates that this system can be used to regulate B. burgdorferi genes in vivo, within the context of an experimental tick-mouse infectious cycle. RpoS is an alternative sigma factor that is required for ospC transcription. However, the role of other temperature-dependent factors has not previously been addressed. Our results with the inducible rpoS strain demonstrate that RpoS alone is sufficient to activate OspC expression, even at 23 degrees C. This is the first functional inducible promoter system developed for use in B. burgdorferi and, for the first time, will provide researchers with the ability to artificially regulate the expression of genes in this pathogenic spirochaete.

Artificial regulation of ospC expression in Borrelia burgdorferi. Publishing Authors By Initials

MA Gilbert,EA Morton,SF Bundle,DS Samuels,

For similar proteins: transcription factors: sigma factor research abstracts see: proteins: transcription factors: sigma factor research

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Artificial regulation of ospC expression in Borrelia burgdorferi. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Molecular microbiology

VOLUME: 63

Page Numbers: 1259-73

Journal Abbreviation: Mol. Microbiol.

ISSN: 0950-382X

DAY: 3

MONTH: Feb

YEAR: 2007

Artificial regulation of ospC expression in Borrelia burgdorferi. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8712028

Artificial regulation of ospC expression in Borrelia burgdorferi. Keywords Mesh Terms:

KEYWORDS: Sigma Factor

MESH TERMS: genetics

Chemical & Substance for Abstract: Artificial regulation of ospC expression in Borrelia burgdorferi. Information

Substance Name: sigma factor KatF protein, Bacteria

Registry Number: 0

Grant and Affiliation Information for Artificial regulation of ospC expression in Borrelia burgdorferi.

AFFILIATION: Division of Biological Sciences, The University of Montana, Missoula, MT 59812-4824, USA.

Country: England

AGENCY: United States NIAID

GRANT: R01 AI051486-05

ACRONYM: AI

MEDLINETA: Mol Microbiol

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