Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡
Identifieur interne : 000491 ( Ncbi/Merge ); précédent : 000490; suivant : 000492Genome-Wide Transposon Mutagenesis in Pathogenic Leptospira Species▿ ‡
Auteurs : Gerald L. Murray ; Viviane Morel ; Gustavo M. Cerqueira ; Julio Croda ; Amporn Srikram ; Rebekah Henry ; Albert I. Ko ; Odir A. Dellagostin ; Dieter M. Bulach ; Rasana W. Sermswan ; Ben Adler ; Mathieu PicardeauSource :
- Infection and Immunity [ 0019-9567 ] ; 2008.
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DOI: 10.1128/IAI.01293-08
PubMed: 19047402
PubMed Central: 2632054
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Genome-Wide Transposon Mutagenesis in Pathogenic <italic>Leptospira</italic> Species<xref ref-type="fn" rid="fn3">▿</xref> <xref ref-type="fn" rid="fn2">‡</xref> </title><author><name sortKey="Murray, Gerald L" sort="Murray, Gerald L" uniqKey="Murray G" first="Gerald L." last="Murray">Gerald L. Murray</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Morel, Viviane" sort="Morel, Viviane" uniqKey="Morel V" first="Viviane" last="Morel">Viviane Morel</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Cerqueira, Gustavo M" sort="Cerqueira, Gustavo M" uniqKey="Cerqueira G" first="Gustavo M." last="Cerqueira">Gustavo M. Cerqueira</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Croda, Julio" sort="Croda, Julio" uniqKey="Croda J" first="Julio" last="Croda">Julio Croda</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Srikram, Amporn" sort="Srikram, Amporn" uniqKey="Srikram A" first="Amporn" last="Srikram">Amporn Srikram</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Henry, Rebekah" sort="Henry, Rebekah" uniqKey="Henry R" first="Rebekah" last="Henry">Rebekah Henry</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ko, Albert I" sort="Ko, Albert I" uniqKey="Ko A" first="Albert I." last="Ko">Albert I. Ko</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Dellagostin, Odir A" sort="Dellagostin, Odir A" uniqKey="Dellagostin O" first="Odir A." last="Dellagostin">Odir A. Dellagostin</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Bulach, Dieter M" sort="Bulach, Dieter M" uniqKey="Bulach D" first="Dieter M." last="Bulach">Dieter M. Bulach</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Sermswan, Rasana W" sort="Sermswan, Rasana W" uniqKey="Sermswan R" first="Rasana W." last="Sermswan">Rasana W. Sermswan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Adler, Ben" sort="Adler, Ben" uniqKey="Adler B" first="Ben" last="Adler">Ben Adler</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Picardeau, Mathieu" sort="Picardeau, Mathieu" uniqKey="Picardeau M" first="Mathieu" last="Picardeau">Mathieu Picardeau</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">19047402</idno><idno type="pmc">2632054</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632054</idno><idno type="RBID">PMC:2632054</idno><idno type="doi">10.1128/IAI.01293-08</idno><date when="2008">2008</date><idno type="wicri:Area/Pmc/Corpus">000F88</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000F88</idno><idno type="wicri:Area/Pmc/Curation">000E49</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000E49</idno><idno type="wicri:Area/Pmc/Checkpoint">002A01</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">002A01</idno><idno type="wicri:Area/Ncbi/Merge">000491</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Genome-Wide Transposon Mutagenesis in Pathogenic <italic>Leptospira</italic> Species<xref ref-type="fn" rid="fn3">▿</xref> <xref ref-type="fn" rid="fn2">‡</xref> </title><author><name sortKey="Murray, Gerald L" sort="Murray, Gerald L" uniqKey="Murray G" first="Gerald L." last="Murray">Gerald L. Murray</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Morel, Viviane" sort="Morel, Viviane" uniqKey="Morel V" first="Viviane" last="Morel">Viviane Morel</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Cerqueira, Gustavo M" sort="Cerqueira, Gustavo M" uniqKey="Cerqueira G" first="Gustavo M." last="Cerqueira">Gustavo M. Cerqueira</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Croda, Julio" sort="Croda, Julio" uniqKey="Croda J" first="Julio" last="Croda">Julio Croda</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Srikram, Amporn" sort="Srikram, Amporn" uniqKey="Srikram A" first="Amporn" last="Srikram">Amporn Srikram</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Henry, Rebekah" sort="Henry, Rebekah" uniqKey="Henry R" first="Rebekah" last="Henry">Rebekah Henry</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ko, Albert I" sort="Ko, Albert I" uniqKey="Ko A" first="Albert I." last="Ko">Albert I. Ko</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Dellagostin, Odir A" sort="Dellagostin, Odir A" uniqKey="Dellagostin O" first="Odir A." last="Dellagostin">Odir A. Dellagostin</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Bulach, Dieter M" sort="Bulach, Dieter M" uniqKey="Bulach D" first="Dieter M." last="Bulach">Dieter M. Bulach</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Sermswan, Rasana W" sort="Sermswan, Rasana W" uniqKey="Sermswan R" first="Rasana W." last="Sermswan">Rasana W. Sermswan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Adler, Ben" sort="Adler, Ben" uniqKey="Adler B" first="Ben" last="Adler">Ben Adler</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Picardeau, Mathieu" sort="Picardeau, Mathieu" uniqKey="Picardeau M" first="Mathieu" last="Picardeau">Mathieu Picardeau</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Infection and Immunity</title><idno type="ISSN">0019-9567</idno><idno type="eISSN">1098-5522</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><italic>Leptospira interrogans</italic> is the most common cause of leptospirosis in humans and animals. Genetic analysis of <italic>L. interrogans</italic> has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the <italic>mariner</italic>-based transposon <italic>Himar1</italic> to generate the first defined mutants in <italic>L. interrogans</italic>. In this study, a total of 929 independent transposon mutants were obtained and the location of insertion determined. Of these mutants, 721 were located in the protein coding regions of 551 different genes. While sequence analysis of transposon insertion sites indicated that transposition occurred in an essentially random fashion in the genome, 25 unique transposon mutants were found to exhibit insertions into genes encoding 16S or 23S rRNAs, suggesting these genes are insertional hot spots in the <italic>L. interrogans</italic> genome. In contrast, loci containing notionally essential genes involved in lipopolysaccharide and heme biosynthesis showed few transposon insertions. The effect of gene disruption on the virulence of a selected set of defined mutants was investigated using the hamster model of leptospirosis. Two attenuated mutants with disruptions in hypothetical genes were identified, thus validating the use of transposon mutagenesis for the identification of novel virulence factors in <italic>L. interrogans</italic>. This library provides a valuable resource for the study of gene function in <italic>L. interrogans</italic>. Combined with the genome sequences of <italic>L. interrogans</italic>, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of <italic>L. interrogans</italic>.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Infect Immun</journal-id><journal-id journal-id-type="publisher-id">iai</journal-id><journal-title>Infection and Immunity</journal-title><issn pub-type="ppub">0019-9567</issn><issn pub-type="epub">1098-5522</issn><publisher><publisher-name>American Society for Microbiology (ASM)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">19047402</article-id><article-id pub-id-type="pmc">2632054</article-id><article-id pub-id-type="publisher-id">1293-08</article-id><article-id pub-id-type="doi">10.1128/IAI.01293-08</article-id><article-categories><subj-group subj-group-type="heading"><subject>Bacterial Infections</subject></subj-group></article-categories><title-group><article-title>Genome-Wide Transposon Mutagenesis in Pathogenic <italic>Leptospira</italic> Species<xref ref-type="fn" rid="fn3">▿</xref> <xref ref-type="fn" rid="fn2">‡</xref> </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Murray</surname><given-names>Gerald L.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Morel</surname><given-names>Viviane</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Cerqueira</surname><given-names>Gustavo M.</given-names></name><xref ref-type="aff" rid="aff1">2</xref><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Croda</surname><given-names>Julio</given-names></name><xref ref-type="aff" rid="aff1">2</xref><xref ref-type="aff" rid="aff1">4</xref></contrib><contrib contrib-type="author"><name><surname>Srikram</surname><given-names>Amporn</given-names></name><xref ref-type="aff" rid="aff1">5</xref></contrib><contrib contrib-type="author"><name><surname>Henry</surname><given-names>Rebekah</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Ko</surname><given-names>Albert I.</given-names></name><xref ref-type="aff" rid="aff1">4</xref><xref ref-type="aff" rid="aff1">6</xref></contrib><contrib contrib-type="author"><name><surname>Dellagostin</surname><given-names>Odir A.</given-names></name><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Bulach</surname><given-names>Dieter M.</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="fn" rid="fn1">†</xref></contrib><contrib contrib-type="author"><name><surname>Sermswan</surname><given-names>Rasana W.</given-names></name><xref ref-type="aff" rid="aff1">7</xref></contrib><contrib contrib-type="author"><name><surname>Adler</surname><given-names>Ben</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="aff" rid="aff1">8</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib><contrib contrib-type="author"><name><surname>Picardeau</surname><given-names>Mathieu</given-names></name><xref ref-type="aff" rid="aff1">2</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Australian Bacterial Pathogenesis Program, Department of Microbiology, Monash University, Clayton, VIC 3800, Australia,<label>1</label> Institut Pasteur, Unité de Biologie des Spirochètes, Paris, France,<label>2</label> Centro de Biotecnologia, Universidade Federal de Pelotas, P.O. Box 354, 96010-900, Pelotas, RS, Brazil,<label>3</label> Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Brazil,<label>4</label> Melioidosis Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand,<label>5</label> Division of International Medicine and Infectious Disease, Weill Medical College of Cornell University, New York, New York,<label>6</label> Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand,<label>7</label> Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Clayton, VIC 3800, Australia<label>8</label></aff><author-notes><fn id="cor1"><label>*</label><p>Mailing address for Mathieu Picardeau: Unité de Biologie des Spirochètes, Institut Pasteur, 28 rue du docteur Roux, 75724 Paris Cedex 15, France. Phone: 33 (1) 45 68 83 68. Fax: 33 (1) 40 61 30 01. E-mail: <email>mpicard@pasteur.fr</email>. Mailing address for Ben Adler: Department of Microbiology, Monash University, Wellington Road, Victoria 3800, Australia. Phone: 61 3 9905 4815. Fax: 61 3 9905 4811. E-mail: <email>ben.adler@med.monash.edu.au</email></p></fn><fn id="fn1"><label>†</label><p>Present address: CSIRO Livestock Industries, Australian Animal Health Laboratory (AAHL), Geelong, VIC 3220, Australia.</p></fn></author-notes><pub-date pub-type="ppub"><month>2</month><year>2009</year></pub-date><pub-date pub-type="epub"><day>1</day><month>12</month><year>2008</year></pub-date><volume>77</volume><issue>2</issue><fpage>810</fpage><lpage>816</lpage><history><date date-type="received"><day>21</day><month>10</month><year>2008</year></date><date date-type="rev-recd"><day>11</day><month>11</month><year>2008</year></date><date date-type="accepted"><day>23</day><month>11</month><year>2008</year></date></history><permissions><copyright-statement>Copyright © 2009, American Society for Microbiology</copyright-statement></permissions><self-uri xlink:title="pdf" xlink:href="zii00209000810.pdf"></self-uri><abstract><p><italic>Leptospira interrogans</italic> is the most common cause of leptospirosis in humans and animals. Genetic analysis of <italic>L. interrogans</italic> has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the <italic>mariner</italic>-based transposon <italic>Himar1</italic> to generate the first defined mutants in <italic>L. interrogans</italic>. In this study, a total of 929 independent transposon mutants were obtained and the location of insertion determined. Of these mutants, 721 were located in the protein coding regions of 551 different genes. While sequence analysis of transposon insertion sites indicated that transposition occurred in an essentially random fashion in the genome, 25 unique transposon mutants were found to exhibit insertions into genes encoding 16S or 23S rRNAs, suggesting these genes are insertional hot spots in the <italic>L. interrogans</italic> genome. In contrast, loci containing notionally essential genes involved in lipopolysaccharide and heme biosynthesis showed few transposon insertions. The effect of gene disruption on the virulence of a selected set of defined mutants was investigated using the hamster model of leptospirosis. Two attenuated mutants with disruptions in hypothetical genes were identified, thus validating the use of transposon mutagenesis for the identification of novel virulence factors in <italic>L. interrogans</italic>. This library provides a valuable resource for the study of gene function in <italic>L. interrogans</italic>. Combined with the genome sequences of <italic>L. interrogans</italic>, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of <italic>L. interrogans</italic>.</p></abstract></article-meta><notes><fn-group><fn><p><italic>Editor:</italic> A. J. Bäumler</p></fn></fn-group></notes></front><floats-wrap><fig position="float" id="f1"><label>FIG. 1.</label><caption><p>Mapping of transposon insertions on the genome of <italic>L. interrogans</italic>. Insertion sites of <italic>Himar1</italic> in 826 transposon mutants (excluding insertions into 16S and 23S rRNA and transposases) of <italic>L. interrogans</italic> were mapped onto circular representations. From the outside in: first, coordinates of the circular chromosome; next, insertion sites of the random mutants in (i) <italic>L. interrogans</italic> serovar Manilae strain L495; (ii) <italic>L. interrogans</italic> serovar Lai strain 56601; (iii) <italic>L. interrogans</italic> serovar Copenhageni strain Fiocruz L1-130; (iv) <italic>L. interrogans</italic> serogroup Canicola strain Kito; (v) <italic>L. interrogans</italic> serovar Pomona strain PO-06-047; and (vi) <italic>L. interrogans</italic> serovar Canicola strain L1-133 (no random mutants in the small chromosome for this strain). Positions of the LPS and <italic>hem</italic> loci are indicated on the large (CI) and small (CII) chromosomes, respectively.</p></caption><graphic xlink:href="zii0020978160001"></graphic></fig><fig position="float" id="f2"><label>FIG. 2.</label><caption><p><italic>Himar1</italic> target site consensus sequence. Sequence logo is drawn from 100 distinct <italic>Himar1</italic> insertion sites in <italic>L. interrogans</italic> serovar Lai strain 56601. The degree of sequence conservation at each position is indicated by the height of letters (maximum of 2 bits for a nucleotide sequence).</p></caption><graphic xlink:href="zii0020978160002"></graphic></fig><table-wrap position="float" id="t1"><label>TABLE 1.</label><caption><p>Bacterial strains used for random transposon mutagenesis</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Species</th><th colspan="1" rowspan="1" align="center" valign="bottom">Serovar<xref ref-type="table-fn" rid="t1fn1"><italic>a</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Strain<xref ref-type="table-fn" rid="t1fn2"><italic>b</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Transformation frequency<xref ref-type="table-fn" rid="t1fn3"><italic>c</italic></xref></th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Copenhageni</td><td colspan="1" rowspan="1" align="left" valign="top">L1 130 LP</td><td colspan="1" rowspan="1" align="center" valign="top">2 × 10<sup>−7</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Copenhageni</td><td colspan="1" rowspan="1" align="left" valign="top">L1 130 HP</td><td colspan="1" rowspan="1" align="center" valign="top">2 × 10<sup>−7</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Lai</td><td colspan="1" rowspan="1" align="left" valign="top">56601 LP</td><td colspan="1" rowspan="1" align="center" valign="top">7 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Lai</td><td colspan="1" rowspan="1" align="left" valign="top">56601 HP</td><td colspan="1" rowspan="1" align="center" valign="top">9 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Icterohaemorrhagiae</td><td colspan="1" rowspan="1" align="left" valign="top">Verdun HP</td><td colspan="1" rowspan="1" align="center" valign="top">1 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Manilae</td><td colspan="1" rowspan="1" align="left" valign="top">L495</td><td colspan="1" rowspan="1" align="center" valign="top">8 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. noguchii</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Autumnalis</td><td colspan="1" rowspan="1" align="left" valign="top">Bonito</td><td colspan="1" rowspan="1" align="center" valign="top">≤1 × 10<sup>−8</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Canicola</td><td colspan="1" rowspan="1" align="left" valign="top">L1 133 LP</td><td colspan="1" rowspan="1" align="center" valign="top">9 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top"><underline>Canicola</underline></td><td colspan="1" rowspan="1" align="left" valign="top">Kito LP</td><td colspan="1" rowspan="1" align="center" valign="top">9 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. interrogans</italic></td><td colspan="1" rowspan="1" align="left" valign="top">Pomona</td><td colspan="1" rowspan="1" align="left" valign="top">PO-06-047</td><td colspan="1" rowspan="1" align="center" valign="top">8 × 10<sup>−6</sup></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"><italic>L. weilii</italic></td><td colspan="1" rowspan="1" align="left" valign="top"><underline>Hebdomadis</underline></td><td colspan="1" rowspan="1" align="left" valign="top">EcoChallenge LP</td><td colspan="1" rowspan="1" align="center" valign="top">5 × 10<sup>−6</sup></td></tr></tbody></table><table-wrap-foot><fn id="t1fn1"><label>a</label><p>When an entry is underlined, only the serogroup of the studied strain is indicated (the serovar was not identified).</p></fn><fn id="t1fn2"><label>b</label><p>LP, low-passage strain; HP, high-passage strain.</p></fn><fn id="t1fn3"><label>c</label><p>Transformation frequency is defined as the number of transposon mutants divided by the number of cells which survived electroporation (approximately 10%). A transformation frequency of ≤1 × 10<sup>−8</sup> represents the limit of detection in these transformations.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t2"><label>TABLE 2.</label><caption><p><italic>L. interrogans</italic> mutant libraries</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Characteristic</th><th colspan="1" rowspan="1" align="center" valign="bottom">Value</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">No. of mutants with defined insertion locations</td><td colspan="1" rowspan="1" align="char" char="." valign="top">929</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">No. of mutants:</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serovar Manilae strain L495</td><td colspan="1" rowspan="1" align="char" char="." valign="top">616</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serovar Lai strain 56601</td><td colspan="1" rowspan="1" align="char" char="." valign="top">249</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serogroup Canicola strain Kito EFS</td><td colspan="1" rowspan="1" align="char" char="." valign="top">32</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serovar Pomona strain EUA</td><td colspan="1" rowspan="1" align="char" char="." valign="top">17</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serovar Copenhageni strain Fiocruz L1-130</td><td colspan="1" rowspan="1" align="char" char="." valign="top">9</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> For <italic>L. interrogans</italic> serovar Canicola strain L1-133</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">No. (%) of mutations:</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In chromosome I (92.4% of genome)<xref ref-type="table-fn" rid="t2fn1"><italic>a</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">826 (90)</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In chromosome II (7.6% of genome)<xref ref-type="table-fn" rid="t2fn1"><italic>a</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">92 (10)</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In coding region</td><td colspan="1" rowspan="1" align="char" char="." valign="top">721<xref ref-type="table-fn" rid="t2fn2"><italic>b</italic></xref></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In transposase</td><td colspan="1" rowspan="1" align="char" char="." valign="top">11<xref ref-type="table-fn" rid="t2fn3"><italic>c</italic></xref></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In rRNA gene</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25<xref ref-type="table-fn" rid="t2fn4"><italic>d</italic></xref></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> In intergenic region</td><td colspan="1" rowspan="1" align="char" char="." valign="top">172</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">No. of ORFs disrupted:</td><td colspan="1" rowspan="1" align="char" char="." valign="top">551</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> Encoding hypothetical proteins</td><td colspan="1" rowspan="1" align="char" char="." valign="top">266</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> Encoding proteins with predicted <italic>L. biflexa</italic> orthologs</td><td colspan="1" rowspan="1" align="char" char="." valign="top">312</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> Encoding proteins with predicted <italic>L. borgpetersenii</italic> orthologs</td><td colspan="1" rowspan="1" align="char" char="." valign="top">437</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> Encoding pathogen-specific proteins</td><td colspan="1" rowspan="1" align="char" char="." valign="top">139</td></tr></tbody></table><table-wrap-foot><fn id="t2fn1"><label>a</label><p>Excluding insertion locations corresponding to multiple locations (transposases).</p></fn><fn id="t2fn2"><label>b</label><p>473 in <italic>L. interrogans</italic> serovar Manilae strain L495, 200 in <italic>L. interrogans</italic> serovar Lai strain 56601, 24 in <italic>L. interrogans</italic> serogroup Canicola strain Kito, 13 in <italic>L. interrogans</italic> serovar Pomona strain PO-06-047, 7 in <italic>L. interrogans</italic> serovar Copenhageni strain Fiocruz L1-130, and 4 in <italic>L. interrogans</italic> serovar Canicola strain L1-133.</p></fn><fn id="t2fn3"><label>c</label><p>Six in <italic>L. interrogans</italic> serovar Lai strain 56601, four in <italic>L. interrogans</italic> serovar Manilae strain L495, and one in <italic>L. interrogans</italic> serogroup Canicola strain Kito.</p></fn><fn id="t2fn4"><label>d</label><p>Nineteen in <italic>L. interrogans</italic> serovar Manilae strain L495, five in <italic>L. interrogans</italic> serovar Lai strain 56601, and one in <italic>L. interrogans</italic> serogroup Canicola strain Kito.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t3"><label>TABLE 3.</label><caption><p>Virulence of mutants in hamster model of acute infection</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Strain<xref ref-type="table-fn" rid="t3fn1"><italic>a</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Location of insertion</th><th colspan="1" rowspan="1" align="center" valign="bottom">Predicted function of mutated gene or description<xref ref-type="table-fn" rid="t3fn2"><italic>b</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Location<xref ref-type="table-fn" rid="t3fn3"><italic>c</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom"><italic>L. biflexa</italic><xref ref-type="table-fn" rid="t3fn4"><italic>d</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Dose(s)<xref ref-type="table-fn" rid="t3fn5"><italic>e</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Hamster survival<xref ref-type="table-fn" rid="t3fn6"><italic>f</italic></xref></th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">L495</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="left" valign="top">Wild-type control</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">EMJH</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="left" valign="top">Negative control</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">5/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M739</td><td colspan="1" rowspan="1" align="center" valign="top">LB125</td><td colspan="1" rowspan="1" align="left" valign="top">Chemotaxis-related protein</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M775*</td><td colspan="1" rowspan="1" align="center" valign="top">LB328</td><td colspan="1" rowspan="1" align="left" valign="top">OmpA family protein</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">2/8</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M776</td><td colspan="1" rowspan="1" align="center" valign="top">LA1857</td><td colspan="1" rowspan="1" align="left" valign="top">Fur paralogue</td><td colspan="1" rowspan="1" align="left" valign="top">C</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/8</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M777</td><td colspan="1" rowspan="1" align="center" valign="top">intergenic</td><td colspan="1" rowspan="1" align="left" valign="top">Intergenic mutant control</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M780</td><td colspan="1" rowspan="1" align="center" valign="top">LA2469</td><td colspan="1" rowspan="1" align="left" valign="top">CheX, inhibitor of MCP methylation</td><td colspan="1" rowspan="1" align="left" valign="top">C</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M789</td><td colspan="1" rowspan="1" align="center" valign="top">LA3028</td><td colspan="1" rowspan="1" align="left" valign="top">HP, contains leucine-rich repeats</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M894</td><td colspan="1" rowspan="1" align="center" valign="top">LA3258</td><td colspan="1" rowspan="1" align="left" valign="top">TonB-dependent receptor</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M880*</td><td colspan="1" rowspan="1" align="center" valign="top">LA4161</td><td colspan="1" rowspan="1" align="left" valign="top">Metalloprotease</td><td colspan="1" rowspan="1" align="left" valign="top">Secreted</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M834</td><td colspan="1" rowspan="1" align="center" valign="top">LA3881</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M895<sup>#</sup></td><td colspan="1" rowspan="1" align="center" valign="top">LA1641</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">IM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup>, 10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">4/4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1061*</td><td colspan="1" rowspan="1" align="center" valign="top">LA0709</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">IM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">1/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M765*</td><td colspan="1" rowspan="1" align="center" valign="top">LA3097</td><td colspan="1" rowspan="1" align="left" valign="top">LipL71</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1224*</td><td colspan="1" rowspan="1" align="center" valign="top">LA1499</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M886*</td><td colspan="1" rowspan="1" align="center" valign="top">LA2003</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1086*</td><td colspan="1" rowspan="1" align="center" valign="top">LA4129</td><td colspan="1" rowspan="1" align="left" valign="top">HP, ankyrin repeat protein</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M874<sup>#</sup></td><td colspan="1" rowspan="1" align="center" valign="top">LA0615</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">5/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M421</td><td colspan="1" rowspan="1" align="center" valign="top">LA4324</td><td colspan="1" rowspan="1" align="left" valign="top">LenE</td><td colspan="1" rowspan="1" align="left" valign="top">OM</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">1/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M911</td><td colspan="1" rowspan="1" align="center" valign="top">LA4209</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>4</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M977</td><td colspan="1" rowspan="1" align="center" valign="top">LA3103</td><td colspan="1" rowspan="1" align="left" valign="top">LenB</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>4</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M983*</td><td colspan="1" rowspan="1" align="center" valign="top">LB362</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1065</td><td colspan="1" rowspan="1" align="center" valign="top">LA1252</td><td colspan="1" rowspan="1" align="left" valign="top">CheB, chemotaxis response regulator</td><td colspan="1" rowspan="1" align="left" valign="top">C</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>4</sup></td><td colspan="1" rowspan="1" align="center" valign="top">2/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1115*</td><td colspan="1" rowspan="1" align="center" valign="top">LA0676</td><td colspan="1" rowspan="1" align="left" valign="top">MCP</td><td colspan="1" rowspan="1" align="left" valign="top">IM</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>5</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1225*</td><td colspan="1" rowspan="1" align="center" valign="top">LA0136</td><td colspan="1" rowspan="1" align="left" valign="top">HP, LipL45-related protein</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1014*</td><td colspan="1" rowspan="1" align="center" valign="top">LA2215</td><td colspan="1" rowspan="1" align="left" valign="top">Flagellar motor protein/OmpA family protein</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">2/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1073*</td><td colspan="1" rowspan="1" align="center" valign="top">LA0137</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">1/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M968</td><td colspan="1" rowspan="1" align="center" valign="top">LA3075</td><td colspan="1" rowspan="1" align="left" valign="top">LigC</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1020*</td><td colspan="1" rowspan="1" align="center" valign="top">LA0505</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1059*</td><td colspan="1" rowspan="1" align="center" valign="top">LA4122</td><td colspan="1" rowspan="1" align="left" valign="top">RNA polymerase sigma subunit</td><td colspan="1" rowspan="1" align="left" valign="top">C</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">M1229*</td><td colspan="1" rowspan="1" align="center" valign="top">LA3367</td><td colspan="1" rowspan="1" align="left" valign="top">HP</td><td colspan="1" rowspan="1" align="left" valign="top">Unknown</td><td colspan="1" rowspan="1" align="center" valign="top">N</td><td colspan="1" rowspan="1" align="left" valign="top">10<sup>3</sup></td><td colspan="1" rowspan="1" align="center" valign="top">0/5</td></tr></tbody></table><table-wrap-foot><fn id="t3fn1"><label>a</label><p>Mutants were constructed in <italic>L. interrogans</italic> serovar Manilae L495. *, the genotype of reisolates was confirmed by PCR; #, bacteria could not be recovered from animals.</p></fn><fn id="t3fn2"><label>b</label><p>Predicted function of protein encoded by disrupted gene. HP, hypothetical protein; MCP, methyl-accepting chemotaxis protein.</p></fn><fn id="t3fn3"><label>c</label><p>Predicted subcellular location of disrupted gene product using the psortb software program (<ext-link ext-link-type="uri" xlink:href="www.psort.org/psortb/">www.psort.org/psortb/</ext-link>); C, cytoplasmic; OM, outer membrane; IM, inner membrane.</p></fn><fn id="t3fn4"><label>d</label><p>Indicates the presence of a deduced protein in <italic>L. biflexa</italic> with >50% similarity by BLASTP. Y, yes; N, no.</p></fn><fn id="t3fn5"><label>e</label><p>Mutants were injected intraperitoneally in two doses (10<sup>3</sup> and 10<sup>5</sup> leptospires) into groups of two or four hamsters, or a single dose was injected into groups of five hamsters.</p></fn><fn id="t3fn6"><label>f</label><p>No. surviving/total. Survival data are pooled for both doses if appropriate.</p></fn></table-wrap-foot></table-wrap></floats-wrap></pmc><affiliations><list></list><tree><noCountry><name sortKey="Adler, Ben" sort="Adler, Ben" uniqKey="Adler B" first="Ben" last="Adler">Ben Adler</name><name sortKey="Bulach, Dieter M" sort="Bulach, Dieter M" uniqKey="Bulach D" first="Dieter M." last="Bulach">Dieter M. Bulach</name><name sortKey="Cerqueira, Gustavo M" sort="Cerqueira, Gustavo M" uniqKey="Cerqueira G" first="Gustavo M." last="Cerqueira">Gustavo M. Cerqueira</name><name sortKey="Croda, Julio" sort="Croda, Julio" uniqKey="Croda J" first="Julio" last="Croda">Julio Croda</name><name sortKey="Dellagostin, Odir A" sort="Dellagostin, Odir A" uniqKey="Dellagostin O" first="Odir A." last="Dellagostin">Odir A. Dellagostin</name><name sortKey="Henry, Rebekah" sort="Henry, Rebekah" uniqKey="Henry R" first="Rebekah" last="Henry">Rebekah Henry</name><name sortKey="Ko, Albert I" sort="Ko, Albert I" uniqKey="Ko A" first="Albert I." last="Ko">Albert I. Ko</name><name sortKey="Morel, Viviane" sort="Morel, Viviane" uniqKey="Morel V" first="Viviane" last="Morel">Viviane Morel</name><name sortKey="Murray, Gerald L" sort="Murray, Gerald L" uniqKey="Murray G" first="Gerald L." last="Murray">Gerald L. Murray</name><name sortKey="Picardeau, Mathieu" sort="Picardeau, Mathieu" uniqKey="Picardeau M" first="Mathieu" last="Picardeau">Mathieu Picardeau</name><name sortKey="Sermswan, Rasana W" sort="Sermswan, Rasana W" uniqKey="Sermswan R" first="Rasana W." last="Sermswan">Rasana W. Sermswan</name><name sortKey="Srikram, Amporn" sort="Srikram, Amporn" uniqKey="Srikram A" first="Amporn" last="Srikram">Amporn Srikram</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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