Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/12331
Title: Genome-wide transposon mutagenesis in pathogenic Leptospira species
Authors: Murray, Gerald L
Morel, Viviane
Cerqueira, Gustavo Maia de
Croda, Julio Henrique Rosa
Srikram, Amporn
Henry, Rebekah
Ko, Albert Icksang
Dellagostin, Odir Antonio
Bulach, Dieter M
Sermswan, Rasana W
Adler, Ben
Picardeau, Mathieu
Affilliation: Monash University. Australian Bacterial Pathogenesis Program. Department of Microbiology. Clayton, Australia
Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France
Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France / Universidade Federal de Pelotas. Centro de Biotecnologia. Pelotas, RS, Brasil
Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Khon Kaen University. Faculty of Medicine. Melioidosis Research Center. Khon Kaen, Thailand
Monash University. Australian Bacterial Pathogenesis Program. Department of Microbiology. Clayton, Australia
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Weill Medical College of Cornell University. Division of International Medicine and Infectious Disease. NY, New York
Universidade Federal de Pelotas. Centro de Biotecnologia. Pelotas, RS, Brasil
Monash University. Australian Bacterial Pathogenesis Program. Department of Microbiology. Clayton, Australia
Khon Kaen University. Faculty of Medicine. Department of Biochemistry. Khon Kaen, Thailand
Monash University. Australian Bacterial Pathogenesis Program. Department of Microbiology. Clayton, Australia / Monash University. Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics. Department of Microbiology. Clayton, Australia
Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France
Abstract: Leptospira interrogans is the most common cause of leptospirosis in humans and animals. Genetic analysis of L. interrogans has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the mariner-based transposon Himar1 to generate the first defined mutants in L. interrogans. In this study, a total of 929 independent transposon mutants were obtained and the location of insertion determined. Of these mutants, 721 were located in the protein coding regions of 551 different genes. While sequence analysis of transposon insertion sites indicated that transposition occurred in an essentially random fashion in the genome, 25 unique transposon mutants were found to exhibit insertions into genes encoding 16S or 23S rRNAs, suggesting these genes are insertional hot spots in the L. interrogans genome. In contrast, loci containing notionally essential genes involved in lipopolysaccharide and heme biosynthesis showed few transposon insertions. The effect of gene disruption on the virulence of a selected set of defined mutants was investigated using the hamster model of leptospirosis. Two attenuated mutants with disruptions in hypothetical genes were identified, thus validating the use of transposon mutagenesis for the identification of novel virulence factors in L. interrogans. This library provides a valuable resource for the study of gene function in L. interrogans. Combined with the genome sequences of L. interrogans, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of L. interrogans.
DeCS: Elementos de DNA Transponíveis/genética
Genoma Bacteriano/genética
Leptospira/fisiologia
Mutagênese Insercional/métodos
Animais
Mapeamento Cromossômico
Cromossomos Bacterianos
Cricetinae
Biblioteca Gênica
Leptospira/genética
Leptospirose/imunologia
Leptospirose/microbiologia
Issue Date: 2009
Publisher: American Society for Microbiology
Citation: MURRAY, G. L. et al. Genome-wide transposon mutagenesis in pathogenic Leptospira species. Infection and Immunity, v. 77, n. 2, p. 810-816, 2009.
DOI: 10.1128/IAI.01293-08
ISSN: 1098-5522
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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