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rdf:type |
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lifeskim:mentions |
umls-concept:C0017262,
umls-concept:C0026336,
umls-concept:C0031669,
umls-concept:C0033268,
umls-concept:C0038172,
umls-concept:C0185117,
umls-concept:C0392756,
umls-concept:C0441655,
umls-concept:C0591833,
umls-concept:C0599946,
umls-concept:C1533691,
umls-concept:C1708096,
umls-concept:C2261241,
umls-concept:C2911684
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pubmed:issue |
1
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pubmed:dateCreated |
1997-2-3
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pubmed:abstractText |
Isogeneic bacterial strains that differ only in the production of a single microbial factor have been invaluable in studying the pathogenesis of bacterial infections. The targeted, intentional inactivation of a gene encoding a potential virulence determinant generally requires homologous recombination to replace the gene with an inactivated allele. To determine whether the insertion and expression of a fragment of a bacterial gene in an antisense orientation could be used as a rapid alternative to allelic inactivation for producing paired isogeneic isolates, we inverted a 600-bp fragment of the Staphylococcus aureus gene encoding alpha-toxin, hla, behind its native promoter on an Escherichia coli-S. aureus shuttle vector. A transformant of an S. aureus strain carrying the antisense hla fragment produced antisense hla RNA and made 16-fold less alpha-toxin than either its parent or an isogeneic transformant containing vector DNA without hla. Also, intraperitoneal injection of 1.5 x 10(9) CFU of the antisense hla-containing transformant was significantly less lethal in a murine model than that of the parent (1 of 10 versus 7 of 10 mice expired [P < 0.02]) or the transformant without hla (1 of 10 versus 7 of 7 mice expired [P < 0.001]). We conclude that the expression of a fragment of hla in an antisense orientation in S. aureus on a plasmid vector reduces alpha-toxin production and the lethal activity of the strain in a murine model. The antisense strategy for creating isogeneic strains of bacteria may facilitate molecular investigations into the pathogenesis of infection. It also may be useful in creating novel live-attenuated strains of bacteria for use as vaccine candidates.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1270140,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1281545,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1652539,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1779933,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1862097,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-1901654,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-2102820,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-2704740,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-2865085,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-2906728,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-3007363,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-3055197,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-3203381,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-3679545,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-5216753,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-6086528,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-6279574,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-6345791,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-6350179,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7541291,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7543479,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7546914,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7601569,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7767645,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7813472,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7817875,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7826024,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7831812,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-7833151,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-8058724,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-8168947,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-8351515,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-8613399,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8975909-942051
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0019-9567
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
65
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
179-84
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8975909-Animals,
pubmed-meshheading:8975909-Bacterial Toxins,
pubmed-meshheading:8975909-Disease Models, Animal,
pubmed-meshheading:8975909-Hemolysin Proteins,
pubmed-meshheading:8975909-Male,
pubmed-meshheading:8975909-Mice,
pubmed-meshheading:8975909-RNA, Antisense,
pubmed-meshheading:8975909-RNA, Bacterial,
pubmed-meshheading:8975909-Staphylococcal Infections,
pubmed-meshheading:8975909-Staphylococcus aureus,
pubmed-meshheading:8975909-Survival Analysis
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pubmed:year |
1997
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pubmed:articleTitle |
Expression of an antisense hla fragment in Staphylococcus aureus reduces alpha-toxin production in vitro and attenuates lethal activity in a murine model.
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pubmed:affiliation |
Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA. kernodds@ctrvax.vanderbilt.edu
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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