000060993 001__ 60993
000060993 005__ 20190709135532.0
000060993 0247_ $$2doi$$a10.3389/fmicb.2017.00315
000060993 0248_ $$2sideral$$a98571
000060993 037__ $$aART-2017-98571
000060993 041__ $$aeng
000060993 100__ $$0(orcid)0000-0003-1027-7583$$aOtal, I.$$uUniversidad de Zaragoza
000060993 245__ $$aDetection of a putative TetR-like gene related to Mycobacterium bovis BCG growth in cholesterol using a gfp-transposon mutagenesis system
000060993 260__ $$c2017
000060993 5060_ $$aAccess copy available to the general public$$fUnrestricted
000060993 5203_ $$aIn vitro transposition is a powerful genetic tool for identifying mycobacterial virulence genes and studying virulence factors in relation to the host. Transposon shuttle mutagenesis is a method for constructing stable insertions in the genome of different microorganisms including mycobacteria. Using an IS1096 derivative, we have constructed the Tngfp, a transposon containing a promoterless green fluorescent protein (gfp) gene. This transposon was able to transpose randomly in Mycobacterium bovis BCG. Bacteria with a single copy of the gfp gene per chromosome from an M. bovis BCG::Tngfp library were analyzed and cells exhibiting high levels of fluorescence were detected by flow cytometry. Application of this approach allowed for the selection of a mutant, BCG_2177c::Tngfp (BCG-Tn), on the basis of high level of long-standing fluorescence at stationary phase. This BCG-Tn mutant showed some particular phenotypic features compared to the wild type strain, mainly during stationary phase, when cholesterol was used as a sole carbon source, thus supporting the relationships of the targeted gene with the regulation of cholesterol metabolism in this bacteria. This approach showed that Tngfp is a potentially useful tool for studying the involvement of the targeted loci in metabolic pathways of mycobacteria.
000060993 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B25$$9info:eu-repo/grantAgreement/ES/MEC/SAB2010-0102$$9info:eu-repo/grantAgreement/ES/MINECO/BIO2014-5258P
000060993 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000060993 590__ $$a4.019$$b2017
000060993 591__ $$aMICROBIOLOGY$$b31 / 125 = 0.248$$c2017$$dQ1$$eT1
000060993 592__ $$a1.699$$b2017
000060993 593__ $$aMicrobiology (medical)$$c2017$$dQ1
000060993 593__ $$aMicrobiology$$c2017$$dQ1
000060993 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000060993 700__ $$aPérez-Herrán, E.
000060993 700__ $$aGarcia-Morales, L.
000060993 700__ $$aMenéndez, M. C.
000060993 700__ $$aGonzalez-y-Merchand, J. A.
000060993 700__ $$0(orcid)0000-0003-2993-5478$$aMartín, C.$$uUniversidad de Zaragoza
000060993 700__ $$aGarcía, M. J.
000060993 7102_ $$11008$$2630$$aUniversidad de Zaragoza$$bDpto. Microb.Med.Pr.,Sal.Públ.$$cÁrea Microbiología
000060993 773__ $$g8 (2017), 315 [13 pp.]$$pFront. microbiol.$$tFRONTIERS IN MICROBIOLOGY$$x1664-302X
000060993 8564_ $$s2114528$$uhttps://zaguan.unizar.es/record/60993/files/texto_completo.pdf$$yVersión publicada
000060993 8564_ $$s22586$$uhttps://zaguan.unizar.es/record/60993/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000060993 909CO $$ooai:zaguan.unizar.es:60993$$particulos$$pdriver
000060993 951__ $$a2019-07-09-12:03:01
000060993 980__ $$aARTICLE