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2012-09-11Zeitschriftenartikel DOI: 10.1186/1471-2180-12-204
Illegitimate recombination: An efficient method for random mutagenesis in Mycobacterium avium subsp. hominissuis
dc.contributor.authorKhattak, Faisal A.
dc.contributor.authorKumar, Ashutosh
dc.contributor.authorKamal, Elisabeth
dc.contributor.authorKunisch, Ralph
dc.contributor.authorLewin, Astrid
dc.date.accessioned2018-05-07T16:08:02Z
dc.date.available2018-05-07T16:08:02Z
dc.date.created2012-12-04
dc.date.issued2012-09-11none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reJLs9D1mcm/PDF/254poAAJwpTd6.pdf
dc.identifier.urihttp://edoc.rki.de/176904/1374
dc.description.abstractBackground: The genus Mycobacterium (M.) comprises highly pathogenic bacteria such as M. tuberculosis as well as environmental opportunistic bacteria called non-tuberculous mycobacteria (NTM). While the incidence of tuberculosis is declining in the developed world, infection rates by NTM are increasing. NTM are ubiquitous and have been isolated from soil, natural water sources, tap water, biofilms, aerosols, dust and sawdust. Lung infections as well as lymphadenitis are most often caused by M. avium subsp. hominissuis (MAH), which is considered to be among the clinically most important NTM. Only few virulence genes from M. avium have been defined among other things due to difficulties in generating M. avium mutants. More efforts in developing new methods for mutagenesis of M. avium and identification of virulence-associated genes are therefore needed. Results: We developed a random mutagenesis method based on illegitimate recombination and integration of a Hygromycin-resistance marker. Screening for mutations possibly affecting virulence was performed by monitoring of pH resistance, colony morphology, cytokine induction in infected macrophages and intracellular persistence. Out of 50 randomly chosen Hygromycin-resistant colonies, four revealed to be affected in virulence-related traits. The mutated genes were MAV_4334 (nitroreductase family protein), MAV_5106 (phosphoenolpyruvate carboxykinase), MAV_1778 (GTP-binding protein LepA) and MAV_3128 (lysyl-tRNA synthetase LysS). Conclusions: We established a random mutagenesis method for MAH that can be easily carried out and combined it with a set of phenotypic screening methods for the identification of virulence-associated mutants. By this method, four new MAH genes were identified that may be involved in virulence.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectVirulenceeng
dc.subjectMutagenesiseng
dc.subjectMycobacteriumeng
dc.subjectMycobacterium avium subsp. hominissuiseng
dc.subjectNon-tuberculous mycobacteriaeng
dc.subjectIllegitimate recombinationeng
dc.subject.ddc610 Medizin
dc.titleIllegitimate recombination: An efficient method for random mutagenesis in Mycobacterium avium subsp. hominissuis
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10028737
dc.identifier.doi10.1186/1471-2180-12-204
dc.identifier.doihttp://dx.doi.org/10.25646/1299
local.edoc.container-titleBMC Microbiology
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.biomedcentral.com/1471-2180/12/204
local.edoc.container-publisher-nameBioMedCentral
local.edoc.container-volume12
local.edoc.container-issue204
local.edoc.container-year2012

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