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We have constructed an inducible promoter system to study the function and regulation of OspC by integrating regulatory elements from the <jats:italic>Escherichia coli lac<\/jats:italic> operon into the <jats:italic>B.\u2003burgdorferi<\/jats:italic> genome. An inducible promoter (<jats:italic>flacp<\/jats:italic>) was constructed by inserting a synthetic <jats:italic>lac<\/jats:italic> operator sequence between the transcriptional start site and the ribosomal binding site of the <jats:italic>B.\u2003burgdorferi flgB<\/jats:italic> promoter; <jats:italic>flacp<\/jats:italic> was then used to replace the native <jats:italic>ospC<\/jats:italic> and <jats:italic>rpoS<\/jats:italic> promoters in <jats:italic>B.\u2003burgdorferi<\/jats:italic> derivatives that constitutively express the <jats:italic>E.\u2003coli<\/jats:italic> Lac repressor protein (LacI). <jats:italic>In vitro<\/jats:italic>, the expression of <jats:italic>ospC<\/jats:italic> and <jats:italic>rpoS<\/jats:italic> from <jats:italic>flacp<\/jats:italic> was dependent on the inducer isopropyl \u03b2\u2010D\u2010thiogalactopyranoside and was unaffected by temperature or pH, conditions commonly used to mimic different aspects of the <jats:italic>B.\u2003burgdorferi<\/jats:italic> 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 <jats:italic>B.\u2003burgdorferi<\/jats:italic> genes <jats:italic>in vivo<\/jats:italic>, within the context of an experimental tick\u2013mouse infectious cycle. RpoS is an alternative sigma factor that is required for <jats:italic>ospC<\/jats:italic> transcription. However, the role of other temperature\u2010dependent factors has not previously been addressed. Our results with the inducible <jats:italic>rpoS<\/jats:italic> strain demonstrate that RpoS alone is sufficient to activate OspC expression, even at 23\u00b0C. This is the first functional inducible promoter system developed for use in <jats:italic>B.\u2003burgdorferi<\/jats:italic> and, for the first time, will provide researchers with the ability to artificially regulate the expression of genes in this pathogenic spirochaete.<\/jats:p>","DOI":"10.1111\/j.1365-2958.2007.05593.x","type":"journal-article","created":{"date-parts":[[2007,1,22]],"date-time":"2007-01-22T18:57:24Z","timestamp":1169492244000},"page":"1259-1273","source":"Crossref","is-referenced-by-count":69,"title":["Artificial regulation of <i>ospC<\/i> expression in <i>Borrelia burgdorferi<\/i>"],"prefix":"10.1111","volume":"63","author":[{"given":"Michael A.","family":"Gilbert","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth A.","family":"Morton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sharyl F.","family":"Bundle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D. 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