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This work was supported by Spanish Ministry of Economy and Competitiveness [grant number BIO2014-5258P]. We also acknowledge the Gobierno de Aragon (DGA-GC B25)/Fondo Social Europeo and Grant CB 2015-255181, CONACyT, Mexico. JG-M held a sabbatical fellowship funded by the Spanish Ministry of Education [SAB2010-0102].

Analysis of institutional authors

Menendez, Maria C.AuthorGarcia, Maria J.Author

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Article

Detection of a Putative TetR-Like Gene Related to Mycobacterium bovis BCG Growth in Cholesterol Using a gfp-Transposon Mutagenesis System

Publicated to:Frontiers in Microbiology. 8 (MAR): 467- - 2017-03-06 8(MAR), DOI: 10.3389/fmicb.2017.00315

Authors: Otal, Isabel; Perez-Herran, Esther; Garcia-Morales, Lazaro; Menendez, Maria C.; Gonzalez-y-Merchand, Jorge A.; Martin, Carlos; Garcia, Maria J.;

Affiliations

Otal, I (reprint author).Univ Zaragoza, Dept Microbiol Med Prevent & Salud Publ, Grp Genet Micobacterias, Zaragoza, Spain. - Author

Abstract

In 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.

Keywords

bcg_2177c genecholesterolgfpmycobacterium bovis bcgtetr-familyAdaptationBcg_2177c geneCholesterolComplexGfpGreen-fluorescent proteinIdentificationIn-vivoMacrophagesMycobacterium bovis bcgPathogenesisReporter transposonsRnaTetr-familyTransposon mutagenesisTuberculosis infection

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Frontiers in Microbiology due to its progression and the good impact it has achieved in recent years, according to the agency Scopus (SJR), it has become a reference in its field. In the year of publication of the work, 2017, it was in position , thus managing to position itself as a Q1 (Primer Cuartil), in the category Microbiology (Medical).

Independientemente del impacto esperado determinado por el canal de difusión, es importante destacar el impacto real observado de la propia aportación.

Según las diferentes agencias de indexación, el número de citas acumuladas por esta publicación hasta la fecha 2025-06-16:

  • WoS: 2
  • Scopus: 2
  • Europe PMC: 1
  • OpenCitations: 2

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-06-16:

  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 22 (PlumX).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: Mexico; United Kingdom.

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: Last Author (GARCIA GARCIA, MARIA JESUS).