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A 2.9-kb fragment containing a putative<jats:italic>spoT<\/jats:italic>gene was isolated from<jats:italic>B. burgdorferi<\/jats:italic>genomic DNA by PCR amplification and cloned into a pBAD24 vector. The cloned gene complemented<jats:italic>Escherichia coli<\/jats:italic>mutant strain CF1693, which contains deletions of both the<jats:italic>relA<\/jats:italic>and<jats:italic>spoT<\/jats:italic>genes. The<jats:italic>spoT<\/jats:italic>gene in<jats:italic>E. coli<\/jats:italic>encodes a bifunctional enzyme capable of synthesizing and degrading (p)ppGpp, which mediates the stringent response during carbon source starvation.<jats:italic>B. burgdorferi<\/jats:italic>has been reported to have a stress response to serum starvation. Thin-layer chromatography was used to detect (p)ppGpp extracted from H<jats:sub>3<\/jats:sub><jats:sup>32<\/jats:sup>PO<jats:sub>4<\/jats:sub>-labeled<jats:italic>B. burgdorferi<\/jats:italic>cells starved for serum in RPMI.<jats:italic>B. burgdorferi spoT<\/jats:italic>gene expression was characterized during fatty acid starvation. Northern analysis of<jats:italic>spoT<\/jats:italic>revealed detectable message at 2.5 min of starvation in RPMI. Expression of<jats:italic>spoT<\/jats:italic>during serum starvation increased \u223c6-fold during the 30 min that starvation conditions were maintained. Further, expression of<jats:italic>spoT<\/jats:italic>decreased when serum was added to serum-starved cells. Reverse transcriptase PCR (RT-PCR) was used to detect<jats:italic>spoT<\/jats:italic>mRNA from \u223c10<jats:sup>6<\/jats:sup>cells starved for serum in RPMI for 2.5 to 30 min or incubated in tick saliva for 15 min. Northern blot analysis suggests that<jats:italic>spoT<\/jats:italic>transcript was \u223c900 nucleotides in length. RT-PCR amplification of the transcript using several sets of primers confirmed this finding. Additionally, a truncated clone containing only the first 950 bp of the 2,001-bp<jats:italic>spoT<\/jats:italic>open reading frame was able to complement<jats:italic>E. coli<\/jats:italic>CF1693. The data suggest that<jats:italic>B. burgdorferi<\/jats:italic>exhibits a stringent response to serum starvation and during incubation in tick saliva.<\/jats:p>","DOI":"10.1128\/jb.185.2.444-452.2003","type":"journal-article","created":{"date-parts":[[2003,1,2]],"date-time":"2003-01-02T19:30:24Z","timestamp":1041535824000},"page":"444-452","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":30,"title":["Expression of<i>spoT<\/i>in<i>Borrelia burgdorferi<\/i>during Serum Starvation"],"prefix":"10.1128","volume":"185","author":[{"given":"Marc B.","family":"Concepcion","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David R.","family":"Nelson","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1099\/00221287-146-1-119","volume":"146","year":"2000","unstructured":"Alban, P. 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