The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis
<p>Abstract</p> <p>Background</p> <p>Plasmids containing <it>hyl</it><sub><it>Efm </it></sub>(pHyl<sub>Efm</sub>) were previously shown to increase gastrointestinal colonization and lethality of <it>Enterococcus faec...
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| Format: | Article |
| Language: | English |
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BMC
2011-01-01
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| Series: | BMC Microbiology |
| Online Access: | http://www.biomedcentral.com/1471-2180/11/20 |
| _version_ | 1811286365903519744 |
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| author | Mojica Maria F Rincón Sandra Montealegre Maria C Panesso Diana Rice Louis B Singh Kavindra V Murray Barbara E Arias Cesar A |
| author_facet | Mojica Maria F Rincón Sandra Montealegre Maria C Panesso Diana Rice Louis B Singh Kavindra V Murray Barbara E Arias Cesar A |
| author_sort | Mojica Maria F |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Plasmids containing <it>hyl</it><sub><it>Efm </it></sub>(pHyl<sub>Efm</sub>) were previously shown to increase gastrointestinal colonization and lethality of <it>Enterococcus faecium </it>in experimental peritonitis. The <it>hyl</it><sub><it>Efm </it></sub>gene, predicting a glycosyl hydrolase, has been considered as a virulence determinant of hospital-associated <it>E. faecium</it>, although its direct contribution to virulence has not been investigated. Here, we constructed mutants of the <it>hyl</it><sub><it>Efm</it></sub>-region and we evaluated their effect on virulence using a murine peritonitis model.</p> <p>Results</p> <p>Five mutants of the <it>hyl</it><sub><it>Efm</it></sub>-region of pHyl<sub>EfmTX16 </sub>from the sequenced endocarditis strain (TX16 [DO]) were obtained using an adaptation of the PheS* system and were evaluated in a commensal strain TX1330RF to which pHyl<sub>EfmTX16 </sub>was transferred by mating; these include <it>i</it>) deletion of <it>hyl</it><sub><it>Efm </it></sub>only; <it>ii</it>) deletion of the gene downstream of <it>hyl</it><sub><it>Efm </it></sub>(<it>down</it>) of unknown function; <it>iii</it>) deletion of <it>hyl</it><sub><it>Efm </it></sub>plus <it>down</it>; <it>iv</it>) deletion of <it>hyl</it><sub><it>Efm</it></sub>-<it>down </it>and two adjacent genes; and <it>v</it>) a 7,534 bp deletion including these four genes plus partial deletion of two others, with replacement by <it>cat</it>. The 7,534 bp deletion did not affect virulence of TX16 in peritonitis but, when pHyl<sub>EfmTX16Δ7,534 </sub>was transferred to the TX1330RF background, the transconjugant was affected in <it>in vitro </it>growth versus TX1330RF(pHyl<sub>EfmTX16</sub>) and was attenuated in virulence; however, neither <it>hyl</it><sub><it>Efm </it></sub>nor <it>hyl</it><sub><it>Efm</it></sub>-<it>down </it>restored wild type function. We did not observe any <it>in vivo </it>effect on virulence of the other deletions of the <it>hyl</it><sub><it>Efm</it></sub>-region</p> <p>Conclusions</p> <p>The four genes of the <it>hyl</it><sub><it>Efm </it></sub>region (including <it>hyl</it><sub><it>Efm</it></sub>) do not mediate the increased virulence conferred by pHyl<sub>EfmTX16 </sub>in murine peritonitis. The use of the markerless counterselection system PheS* should facilitate the genetic manipulation of <it>E. faecium </it>in the future.</p> |
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| format | Article |
| id | doaj.art-62050f27aa12463bb9d6a9861aa6c6b1 |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-04-13T02:58:32Z |
| publishDate | 2011-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-62050f27aa12463bb9d6a9861aa6c6b12022-12-22T03:05:30ZengBMCBMC Microbiology1471-21802011-01-011112010.1186/1471-2180-11-20The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitisMojica Maria FRincón SandraMontealegre Maria CPanesso DianaRice Louis BSingh Kavindra VMurray Barbara EArias Cesar A<p>Abstract</p> <p>Background</p> <p>Plasmids containing <it>hyl</it><sub><it>Efm </it></sub>(pHyl<sub>Efm</sub>) were previously shown to increase gastrointestinal colonization and lethality of <it>Enterococcus faecium </it>in experimental peritonitis. The <it>hyl</it><sub><it>Efm </it></sub>gene, predicting a glycosyl hydrolase, has been considered as a virulence determinant of hospital-associated <it>E. faecium</it>, although its direct contribution to virulence has not been investigated. Here, we constructed mutants of the <it>hyl</it><sub><it>Efm</it></sub>-region and we evaluated their effect on virulence using a murine peritonitis model.</p> <p>Results</p> <p>Five mutants of the <it>hyl</it><sub><it>Efm</it></sub>-region of pHyl<sub>EfmTX16 </sub>from the sequenced endocarditis strain (TX16 [DO]) were obtained using an adaptation of the PheS* system and were evaluated in a commensal strain TX1330RF to which pHyl<sub>EfmTX16 </sub>was transferred by mating; these include <it>i</it>) deletion of <it>hyl</it><sub><it>Efm </it></sub>only; <it>ii</it>) deletion of the gene downstream of <it>hyl</it><sub><it>Efm </it></sub>(<it>down</it>) of unknown function; <it>iii</it>) deletion of <it>hyl</it><sub><it>Efm </it></sub>plus <it>down</it>; <it>iv</it>) deletion of <it>hyl</it><sub><it>Efm</it></sub>-<it>down </it>and two adjacent genes; and <it>v</it>) a 7,534 bp deletion including these four genes plus partial deletion of two others, with replacement by <it>cat</it>. The 7,534 bp deletion did not affect virulence of TX16 in peritonitis but, when pHyl<sub>EfmTX16Δ7,534 </sub>was transferred to the TX1330RF background, the transconjugant was affected in <it>in vitro </it>growth versus TX1330RF(pHyl<sub>EfmTX16</sub>) and was attenuated in virulence; however, neither <it>hyl</it><sub><it>Efm </it></sub>nor <it>hyl</it><sub><it>Efm</it></sub>-<it>down </it>restored wild type function. We did not observe any <it>in vivo </it>effect on virulence of the other deletions of the <it>hyl</it><sub><it>Efm</it></sub>-region</p> <p>Conclusions</p> <p>The four genes of the <it>hyl</it><sub><it>Efm </it></sub>region (including <it>hyl</it><sub><it>Efm</it></sub>) do not mediate the increased virulence conferred by pHyl<sub>EfmTX16 </sub>in murine peritonitis. The use of the markerless counterselection system PheS* should facilitate the genetic manipulation of <it>E. faecium </it>in the future.</p>http://www.biomedcentral.com/1471-2180/11/20 |
| spellingShingle | Mojica Maria F Rincón Sandra Montealegre Maria C Panesso Diana Rice Louis B Singh Kavindra V Murray Barbara E Arias Cesar A The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis BMC Microbiology |
| title | The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis |
| title_full | The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis |
| title_fullStr | The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis |
| title_full_unstemmed | The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis |
| title_short | The <it>hyl</it><sub><it>Efm </it></sub>gene in pHyl<sub>Efm </sub>of <it>Enterococcus faecium </it>is not required in pathogenesis of murine peritonitis |
| title_sort | it hyl it sub it efm it sub gene in phyl sub efm sub of it enterococcus faecium it is not required in pathogenesis of murine peritonitis |
| url | http://www.biomedcentral.com/1471-2180/11/20 |
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