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dc.contributor.authorFeltwell, Theresaen
dc.contributor.authorDorman, Matthewen
dc.contributor.authorGoulding, David Aen
dc.contributor.authorParkhill, Julianen
dc.contributor.authorShort, Francesca Len
dc.date.accessioned2019-09-04T23:31:15Z
dc.date.available2019-09-04T23:31:15Z
dc.date.issued2019-01-07en
dc.identifier.issn1940-087X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/296435
dc.description.abstractCapsule is a key virulence factor in many bacterial species, mediating immune evasion and resistance to various physical stresses. While many methods are available to quantify and compare capsule production between different strains or mutants, there is no widely used method for sorting bacteria based on how much capsule they produce. We have developed a method to separate bacteria by capsule amount, using a discontinuous density gradient. This method is used to compare capsule amounts semi-quantitatively between cultures, to isolate mutants with altered capsule production, and to purify capsulated bacteria from complex samples. This method can also be coupled with transposon-insertion sequencing to identify genes involved in capsule regulation. Here, the method is demonstrated in detail, including how to optimize the gradient conditions for a new bacterial species or strain, and how to construct and run the density gradient.
dc.format.mediumElectronicen
dc.languageengen
dc.publisherMYJoVE Corporation
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectAnimalsen
dc.subjectBacterial Capsulesen
dc.subjectVirulence Factorsen
dc.titleSeparating Bacteria by Capsule Amount Using a Discontinuous Density Gradient.en
dc.typeArticle
prism.issueIdentifier143en
prism.publicationDate2019en
prism.publicationNameJournal of visualized experiments : JoVEen
dc.identifier.doi10.17863/CAM.43485
dcterms.dateAccepted2018-12-03en
rioxxterms.versionofrecord10.3791/58679en
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2019-01-07en
dc.contributor.orcidDorman, Matthew [0000-0001-7064-6163]
dc.contributor.orcidParkhill, Julian [0000-0002-7069-5958]
dc.contributor.orcidShort, Francesca L [0000-0002-0025-4858]
dc.identifier.eissn1940-087X
rioxxterms.typeJournal Article/Reviewen


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International