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Toolbox of Molecular Techniques for Studying Leptospira Spp.

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Spirochete Biology: The Post Genomic Era

Part of the book series: Current Topics in Microbiology and Immunology ((CT MICROBIOLOGY,volume 415))

Abstract

This chapter covers the progress made in the Leptospira field since the application of mutagenesis techniques and how they have allowed the study of virulence factors and, more generally, the biology of Leptospira. The last decade has seen advances in our ability to perform molecular genetic analysis of Leptospira. Major achievements include the generation of large collections of mutant strains and the construction of replicative plasmids, enabling complementation of mutations. However, there are still no practical tools for routine genetic manipulation of pathogenic Leptospira strains, slowing down advances in pathogenesis research. This review summarizes the status of the molecular genetic toolbox for Leptospira species and highlights new challenges in the nascent field of Leptospira genetics.

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References

  • Adler B, Lo M, Seemann T, Murray GL (2011) Pathogenesis of leptospirosis: the influence of genomics. Vet Microbiol 153:73–81

    Article  PubMed  CAS  Google Scholar 

  • Aviat F, Slamti L, Cerqueira GM, Lourdault K, Picardeau M (2010) Expanding the genetic toolbox for Leptospira species by generation of fluorescent bacteria. Appl Environ Microbiol 76:8135–8142

    Google Scholar 

  • Baril C, Richaud C, Fournie E, Baranton G, Saint Girons I (1992) Cloning of dapD, aroD and asd of Leptospira interrogans serovar icterohaemorrhagiae, and nucleotide sequence of the asd gene. J Gen Microbiol 138:47–53

    Article  PubMed  CAS  Google Scholar 

  • Bauby H, Saint Girons I, Picardeau M (2003) Construction and complementation of the first auxotrophic mutant in the spirochaete Leptospira meyeri. Microbiology 149:689–693

    Article  PubMed  CAS  Google Scholar 

  • Beck M, Malmström JA, Lange V, Schmidt A, Deutsch EW, Aebersold R (2009) Visual proteomics of the human pathogen Leptospira interrogans. Nat Methods 6:817–823

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Belfaiza J, Martel A, Margarita D, Saint Girons I (1998) Direct sulfhydrylation for methionine biosynthesis in Leptospira meyeri. J Bacteriol 180:250–255

    PubMed  PubMed Central  CAS  Google Scholar 

  • Bono JL, Elias AF, Kupko JJ, Stevenson B, Tilly K, Rosa P (2000) Efficient targeted mutagenesis in Borrelia burgdorferi. J Bacteriol 182:2445–2452

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Bourhy P, Frangeul L, Couve E, Glaser P, Saint Girons I, Picardeau M (2005a) Complete nucleotide sequence of the LE1 prophage from the spirochete Leptospira biflexa and characterization of its replication and partition functions. J Bacteriol 187:3931–3940

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Bourhy P, Louvel H, Saint Girons I, Picardeau M (2005b) Random insertional mutagenesis of Leptospira interrogans, the agent of leptospirosis, using a mariner transposon. J Bacteriol 187:3255–3258

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Bourhy P, Salaün L, Lajus A, Médigue C, Boursaux-Eude C, Picardeau M (2007) A genomic island of the pathogen Leptospira interrogans serovar Lai can excise from its chromosome. Infect Immun 75:677–683

    Article  PubMed  CAS  Google Scholar 

  • Bromley DB, Charon NW (1979) Axial filament involvement in the motility of Leptospira interrogans. J Bacteriol 137:1406–1412

    PubMed  PubMed Central  CAS  Google Scholar 

  • Bulach DM, Zuerner RL, Wilson P, Seemann T, McGrath A, Cullen PA, Davis J, Johnson M, Kuczek E, Alt DP, Peterson-Burch B, Coppel RL, Rood JI, Davies JK, Adler B (2006) Genome reduction in Leptospira borgpetersenii reflects limited transmission potential. Proc Natl Acad Sci U S A 103:14560–14565

    Article  PubMed  PubMed Central  Google Scholar 

  • Caimano MJ, Sivasankaran SK, Allard A, Hurley D, Hokamp K, Grassmann AA, Hinton JC, Nally JE (2014) A model system for studying the transcriptomic and physiological changes associated with mammalian host-adaptation by Leptospira interrogans serovar Copenhageni. PLoS Pathog 10:e1004004

    Article  PubMed  PubMed Central  Google Scholar 

  • Cao XJ, Dai J, Xu H, Nie S, Chang X, Hu BY, Sheng QH, Wang LS, Ning ZB, Li YX, Guo XK, Zhao GP, Zeng R (2010) High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans. Cell Res 20:197–210

    Article  PubMed  CAS  Google Scholar 

  • Castiblanco-Valencia MM, Fraga TR, Breda LC, Vasconcellos SA, Figueira CP, Picardeau M, Wunder E, Ko AI, Barbosa AS, Isaac L (2016) Acquisition of negative complement regulators by the saprophyte Leptospira biflexa expressing LigA or LigB confers enhanced survival in human serum. Immunol Lett 173:61–68

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cerqueira GM, Souza NM, Araújo ER, Barros AT, Morais ZM, Vasconcellos SA, Nascimento AL (2011) Development of transcriptional fusions to assess Leptospira interrogans promoter activity. PLoS ONE 6:e17409

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Choy HA, Kelley MM, Croda J, Matsunaga J, Babbitt JT, Ko AI, Picardeau M, Haake DA (2011) The multifunctional LigB adhesin binds homeostatic proteins with potential roles in cutaneous infection by pathogenic Leptospira interrogans. PLoS One 6(2):e16879

    Google Scholar 

  • Croda J, Figueira CP, Wunder EAJ, Santos CS, Reis MG, Ko AI, Picardeau M (2008) Targeted mutagenesis in pathogenic Leptospira: disruption of the ligB gene does not affect virulence in animal models of leptospirosis. Infect Immun 76:5826–5833

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cullen PA, Cordwell SJ, Bulach DM, Haake DA, Adler B (2002) Global analysis of outer membrane proteins from Leptospira interrogans serovar Lai. Infect Immun 70:2311–2318

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cullen PA, Haake DA, Adler B (2004) Outer membrane proteins of pathogenic spirochetes. FEMS Microbiol Rev 28:291–318

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cullen PA, Xu X, Matsunaga J, Sanchez Y, Ko AI, Haake DA, Adler B (2005) Surfaceome of Leptospira spp. Infect Immun 73:4853–4863

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Eshghi A, Cullen PA, Cowen L, Zuerner RL, Cameron CE (2009) Global proteome analysis of Leptospira interrogans. J Proteome Res 8:4564–4578

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Eshghi A, Lourdault K, Murray GL, Bartpho T, Sermswan RW, Picardeau M, Adler B, Snarr B, Zuerner RL, Cameron CE (2012a) Leptospira interrogans catalase is required for resistance to H2O2 and for virulence. Infect Imm 80:3892–3899

    Article  CAS  Google Scholar 

  • Eshghi A, Pinne M, Haake DA, Zuerner RL, Frank A, Cameron CE (2012b) Methylation and in vivo expression of the surface-exposed Leptospira interrogans outer-membrane protein OmpL32. Microbiology 158:622–635

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Eshghi A, Becam J, Lambert A, Sismeiro O, Dillies MA, Jagla B, Wunder EA Jr, Ko AI, Coppee JY, Goarant C, Picardeau M (2014) A putative regulatory genetic locus modulates virulence in the pathogen Leptospira interrogans. Infect Immun 82:2542–2552

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Eshghi A, Henderson J, Trent MS, Picardeau M (2015a) Leptospira interrogans lpxD homologue is required for thermal acclimatization and virulence. Infect Immun 83:4314–4321

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Eshghi A, Pappalardo E, Hester S, Thomas B, Pretre G, Picardeau M (2015b) Pathogenic Leptospira interrogans exoproteins are primarily involved in heterotrophic processes. Infect Imm 83:3061–3073

    Article  CAS  Google Scholar 

  • Falkow S (1988) Molecular Koch’s postulates applied to microbial pathogenicity. Rev Infect Dis 10:274–276

    Article  Google Scholar 

  • Figueira CP, Croda J, Choy HA, Haake DA, Reis MG, Ko AI, Picardeau M (2011) Heterologous expression of pathogen-specific genes ligA and ligB in the saprophyte Leptospira biflexa confers enhanced adhesion to cultured cells and fibronectin. BMC Microbiol 11:129

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Fontana C, Lambert A, Benaroudj N, Gasparini D, Gorgette O, Cachet N, Bomchil N, Picardeau M (2016) Analysis of a spontaneous non-motile and avirulent mutant shows that FliM is Required for full endoflagella assembly in Leptospira interrogans. PLoS ONE 11:e0152916

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Fouts DE, Matthias MA, Adhikarla H, Adler B, Berg DE, Bulach D, Buschiazzo A, Chang Y-F, Galloway RL, Haake DA, Haft DH, Hartskeerl R, Ko A, Levett P, Matsunaga J, Mechaly AE, Monk J, Nascimento A, Nelson KE, Palsson A, Peacock SJ, Picardeau M, Ricaldi JN, Thaipandungpanit J, Wunder E, Yang F, Zhang JJ, Vinetz JM (2016) What makes a bacterial species pathogenic?: Comparative genomic analysis of the genus Leptospira. PLoS Negl Trop Dis 10:e0004403

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Gamberini M, Gómez RM, Atzingen MV, Martins EA, Vasconcellos SA, Romero EC, Leite LC, Ho PL, Nascimento AL (2005) Whole-genome analysis of Leptospira interrogans to identify potential vaccine candidates against leptospirosis. FEMS Microbiol Lett 244(2):305–313

    Google Scholar 

  • Haake DA, Martinich C, Summers TA, Shang ES, Pruetz JD, McCoy AM, Mazel MK, Bolin CA (1998) Characterization of leptospiral outer membrane lipoprotein LipL36: downregulation associated with late-log-phase growth and mammalian infection. Infect Immun 66:1579–1587

    PubMed  PubMed Central  CAS  Google Scholar 

  • Haake DA, Matsunaga J (2010) Leptospira: a spirochaete with a hybrid outer membrane. Mol Microbiol 77:805–814

    PubMed  PubMed Central  CAS  Google Scholar 

  • Henry R, Lo M, Khoo C, Zhang H, Boysen RI, Picardeau M, Murray GL, Bulach DM, Adler B, print] MEao (2013). Iron precipitation on the surface of Leptospira interrogans is associated with mutation of the stress-response metalloprotease HtpX. Appl Environ Microbiol 79:4653–4660

    Google Scholar 

  • Kassegne K, Hu W, Ojcius DM, Sun D, Ge Y, Zhao J, Yang XF, Li L, Yan J (2014) Identification of collagenase as a critical virulence factor for invasiveness and transmission of pathogenic leptospira species. J Infect Dis 209:1105–1115

    Article  PubMed  CAS  Google Scholar 

  • Kawabata H, Norris SJ, Watanabe H (2004) BBE02 disruption mutants of Borrelia burgdorferi B31 have a highly transformable, infectious phenotype. Infect Imm 72:7147–7154

    Article  CAS  Google Scholar 

  • King AM, Bartpho T, Sermswan RW, Bulach DM, Eshghi A, Picardeau M, Adler B, Murray GL (2013) Leptospiral outer membrane protein LipL41 is not essential for acute leptospirosis, but requires a small chaperone, Lep, for stable expression. Infect Immun 81:2768–2776

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • King AM, Pretre G, Bartpho T, Sermswan RW, Toma C, Suzuki T, Eshghi A, Picardeau M, Adler B, Murray GL (2014) High-temperature protein G is an essential virulence factor of Leptospira interrogans. Infect Immun 82:1123–1131

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Komi KK, Ge YM, Xin XY, Ojcius DM, Sun D, Hu WL, Zhao X, Lin X, Yan J (2015) ChpK and MazF of the toxin-antitoxin modules are involved in the virulence of Leptospira interrogans during infection. Microbes Infect 17:34–37

    Article  PubMed  CAS  Google Scholar 

  • Lambert A, Picardeau M, Haake DA, Sermswan RW, Srikram A, Adler B, Murray GA (2012) FlaA proteins in Leptospira interrogans are essential for motility and virulence but are not required for formation of the flagellum sheath. Infect Immun 80:2019–2025

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lehmann JS, Fouts DE, Haft DH, Cannella AP, Ricaldi JN, Brinkac L, Harkins D, Durkin S, Sanka R, Sutton G, Moreno A, Vinetz JM, Matthias MA (2013) Pathogenomic inference of virulence-associated genes in Leptospira interrogans. PLoS Negl Trop Dis 7:e2468

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lehmann JS, Corey VC, Ricaldi JN, Vinetz JM, Winzeler EA, Matthias MA (2016) Whole genome shotgun sequencing shows selection on Leptospira regulatory proteins during in vitro culture attenuation. Am J Trop Med Hyg 94:302–313

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lessa-Aquino C, Wunder EA Jr, Lindow JC, Rodrigues CB, Pablo J, Nakajima R, Jasinskas A, Liang L, Reis MG, Ko AI, Medeiros MA, Felgner PL (2015) Proteomic features predict seroreactivity against leptospiral antigens in leptospirosis patients. J Proteome Res 14:549–556

    Article  PubMed  CAS  Google Scholar 

  • Liang L, Felgner PL (2015) A systems biology approach for diagnostic and vaccine antigen discovery in tropical infectious diseases. Curr Opin Infect Dis 28:438–445

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Liao S, Sun A, Ojcius DM, Wu S, Zhao J, Yan J (2009) Inactivation of the fliY gene encoding a flagellar motor switch protein attenuates mobility and virulence of Leptospira interrogans strain Lai. BMC Microbiol 9:253

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lo M, Bulach DM, Powell DR, Haake DA, Matsunaga J, Paustian ML, Zuerner RL, Adler B (2006) Effects of temperature on gene expression patterns in Leptospira interrogans serovar Lai as assessed by whole-genome microarrays. Infect Immun 74:848–859

    Article  CAS  Google Scholar 

  • Lo M, Cordwell SJ, Bulach DM, Adler B (2009) Comparative transcriptional and translational analysis of leptospiral outer membrane protein expression in response to temperature. PLoS Negl Trop Dis 3:e560

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lo M, Murray GL, Khoo CA, Haake DA, Zuerner RL, Adler B (2010) Transcriptional response of Leptospira interrogans to iron limitation and characterization of a PerR homolog. Infect Immun 78:4850–4859

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lourdault K, Cerqueira GM, Wunder EA, Picardeau M (2011). Inactivation of clpB in the pathogen Leptospira interrogans reduces virulence and resistance to stress conditions. Infect Immun 79:3711–3717

    Google Scholar 

  • Lourdault K, Matsunaga J, Haake DA (2016) High-Throughput parallel sequencing to measure fitness of Leptospira interrogans transposon insertion mutants during acute infection. PLoS Negl Trop Dis 10:e0005117

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Louvel H, Saint Girons I, Picardeau M (2005) Isolation and characterization of FecA- and FeoB-mediated iron acquisition systems of the spirochete Leptospira biflexa by random insertional mutagenesis. J Bacteriol 187:3249–3254

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Louvel H, Bommezzadri S, Zidane N, Boursaux-Eude C, Creno S, Magnier A, Rouy Z, Medigue C, Girons IS, Bouchier C, Picardeau M (2006) Comparative and functional genomic analyses of iron transport and regulation in Leptospira spp. J Bacteriol 188:7893–7904

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Louvel H, Picardeau M (2007) Genetic manipulation of Leptospira biflexa. Wiley, Hoboken

    Google Scholar 

  • Louvel H, Betton JM, Picardeau M (2008) Heme rescues a two-component system Leptospira biflexa mutant. BMC Microbiol 8:25

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Malmström J, Beck M, Schmidt A, Lange V, Deutsch EW, Aebersold R (2009) Proteome-wide cellular protein concentrations of the human pathogen Leptospira interrogans. Nature 460:762–765

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Marcsisin RA, Bartpho T, Bulach DM, Srikram A, Sermswan RW, Adler B, Murray GL (2013) Use of a high-throughput screen to identify Leptospira mutants unable to colonize the carrier host or cause disease in the acute model of infection. J Med Microbiol 62:1601–1608

    Article  PubMed  CAS  Google Scholar 

  • Matsunaga J, Coutinho ML (2012) Positive regulation of Leptospira interrogans kdp expression by KdpE as demonstrated with a novel β-galactosidase reporter in Leptospira biflexa. Appl Environ Microbiol 78:5699–5707

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Matsunaga J, Barocchi MA, Croda J, Young TA, Sanchez Y, Siqueira I, Bolin CA, Reis MG, Riley LW, Haake DA, Ko AI (2003) Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. Mol Microbiol 49:929–945

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Matsunaga J, Lo M, Bulach DM, Zuerner RL, Adler B, Haake DA (2007) Response of Leptospira interrogans to physiologic osmolarity: relevance in signaling the environment-to-host transition. Infect Immun 75:2864–2874

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Monahan AM, Callanan JJ, Nally JE (2008) Proteomic analysis of Leptospira interrogans shed in urine of chronically infected hosts. Infect Immun 76:4952–4958

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Murray GL, Morel V, Cerqueira GM, Croda J, Srikram A, Henry R, Ko AI, Dellagostin OA, Bulach DM, Sermswan R, Adler B, Picardeau M (2009a) Genome-wide transposon mutagenesis in pathogenic Leptospira spp. Infect Immun 77:810–816

    Article  PubMed  CAS  Google Scholar 

  • Murray GL, Srikram A, Hoke DE, Wunder EAJ, Henry R, Lo M, Zhang K, Sermswan RW, Ko AI, Adler B (2009b) The major surface protein LipL32 is not required for either acute or chronic infection with Leptospira interrogans. Infect Immun 77:952–958

    Article  PubMed  CAS  Google Scholar 

  • Murray GL, King AM, Srikram A, Sermswan RW, Adler B (2010) Use of luminescent Leptospira interrogans for enumeration in biological assays. J Clin Microbiol 48:2037–2042

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Nally JE, Timoney JF, Stevenson B (2001) Temperature-regulated protein synthesis by Leptospira interrogans. Infect Immun 69:400–404

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Nally JE, Chow E, Fishbein MC, Blanco DR, Lovett MA (2005a) Changes in lipopolysaccharide O antigen distinguish acute versus chronic Leptospira interrogans infections. Infect Immun 73:3251–3260

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Nally JE, Whitelegge JP, Aguilera R, Pereira MM, Blanco DR, Lovett MA (2005b) Purification and proteomic analysis of outer membrane vesicles from a clinical isolate of Leptospira interrogans serovar Copenhageni. Proteomics 5:144–152

    Article  PubMed  CAS  Google Scholar 

  • Nally JE, Whitelegge JP, Bassilian S, Blanco DR, Lovett MA (2007) Characterization of the outer membrane proteome of Leptospira interrogans expressed during acute lethal infection. Infect Immun 75:766–773

    Article  PubMed  CAS  Google Scholar 

  • Nally JE, Monahan AM, Miller IS, Bonilla-Santiago R, Souda P, Whitelegge JP (2011) Comparative proteomic analysis of differentially expressed proteins in the urine of reservoir hosts of leptospirosis. PLoS ONE 6:e26046

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Nally JE, Mullen W, Callanan JJ, Mischak H, Albalat A (2015) Detection of urinary biomarkers in reservoir hosts of leptospirosis by capillary electrophoresis-mass spectrometry. Proteomics Clin Appl 9:543–551

    Article  PubMed  CAS  Google Scholar 

  • Narayanavari SA, Lourdault K, Sritharan M, Haake DA, Matsunaga J (2015) Role of sph2 gene regulation in hemolytic and sphingomyelinase activities produced by Leptospira interrogans. PLoS Negl Trop Dis 9:e0003952

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ozuru R, Saito M, Kanemaru T, Miyahara S, Villanueva SY, Murray GL, Adler B, Fujii J, Yoshida SI (2017) Adipose tissue is the first colonization site of Leptospira interrogans in subcutaneously infected hamsters. PLoS ONE 12:e0172973

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pappas CJ, Picardeau M (2015) Control of gene expression in Leptospira spp. by transcription activator-like effectors (TALEs) demonstrates a potential role for LigA and LigB in virulence in L. interrogans. Appl Environ Microbiol 81:7888–7892

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pappas CJ, Benaroudj N, Picardeau M (2015) A replicative plasmid vector allows efficient complementation of pathogenic Leptospira strains. Appl Environ Microbiol 81:3176–3781

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Patarakul K, Lo M, Adler B (2010) Global transcriptomic response of Leptospira interrogans serovar Copenhageni upon exposure to serum. BMC Microbiol 10:31

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • PÄ›troÅ¡ová H, Picardeau M (2014) Screening of a Leptospira biflexa mutant library to identify genes involved in ethidium bromide tolerance. Appl Environ Microbiol 80:6091–6103

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Picardeau M (2008) Conjugative transfer between Escherichia coli and Leptospira spp. as a new genetic tool. Appl Environ Microbiol 74:319–322

    Article  PubMed  CAS  Google Scholar 

  • Picardeau M (2014) Family Leptospiraceae. In: Rosenberg E (ed) The prokaryotes. Springer, Berlin, pp 711–729

    Google Scholar 

  • Picardeau M, Brenot A, Saint Girons I (2001) First evidence for gene replacement in Leptospira spp. Inactivation of L. biflexa flaB results in non-motile mutants deficient in endoflagella. Mol Microbiol 40:189–199

    Article  PubMed  CAS  Google Scholar 

  • Picardeau M, Bulach DM, Bouchier C, Zuerner RL, Zidane N, Wilson PJ, Creno S, Kuczek ES, Bommezzadri S, Davis JC, McGrath A, Johnson MJ, Boursaux-Eude C, Seemann T, Rouy Z, Coppel RL, Rood JI, Lajus A, Davies JK, Médigue C, Adler B (2008) Genome sequence of the saprophyte Leptospira biflexa provides insights into the evolution of Leptospira and the pathogenesis of leptospirosis. PLoS ONE 3:e1607

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pinne M, Matsunaga J, Haake DA (2012) Leptospiral outer membrane protein microarray, a novel approach to identification of host ligand-binding proteins. J Bacteriol 194:6074–6087

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pinne M, Haake DA (2013) LipL32 Is a Subsurface Lipoprotein of Leptospira interrogans: presentation of new data and reevaluation of previous studies. PLoS ONE 8:e51025

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Poggi D, Oliveira de Giuseppe P, Picardeau M (2010) Antibiotic resistance markers for genetic manipulations of Leptospira spp. Appl Environ Microbiol 76:4882–4885

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Qin JH, Sheng YY, Zhang ZM, Shi YZ, He P, Hu BY, Yang Y, Liu SG, Zhao GP, Guo XK (2006) Genome-wide transcriptional analysis of temperature shift in Leptospira. interrogans serovar lai strain 5660. BMC Microbiol 6:51

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ratet G, Veyrier FJ, Fanton d’Andon M, Kammerscheit X, Nicola MA, Picardeau M, Boneca IG, Werts C (2014) Live imaging of bioluminescent Leptospira interrogans in mice reveals renal colonization as a stealth escape from the blood defenses and antibiotics. PLoS Negl Trop Dis 8:e3359

    Article  PubMed  PubMed Central  Google Scholar 

  • Ren S, Fu G, Jiang X, Zeng R, Xiong H, Lu G, Jiang HQ, Miao Y, Xu H, Zhang Y, Guo X, Shen Y, Qiang BQ, Xia Q, Danchin A, SaintGirons I, Somerville RL, Weng YM, Shi M, Chen Z, Xu JG, Zhao GP (2003) Unique and physiological and pathogenic features of Leptospira interrogans revealed by whole genome sequencing. Nature 422:888–893

    Article  PubMed  CAS  Google Scholar 

  • Ricaldi JN, Fouts DE, Selengut JD, Harkins DM, Moreno A, Lehmann JS, Purushe J, Sanka R, Torres M, Webster NJ, Vinetz JM, Matthias MA (2012a) Whole genome analysis of Leptospira licerasiae provides insight into Leptospiral evolution and pathogenicity. PLoS Negl Trop Dis 6:e1853

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ricaldi JN, Matthias MA, Vinetz JM, Lewis AL (2012b) Expression of sialic acids and other nonulosonic acids in Leptospira. BMC Microbiol 12:161

    Article  PubMed  CAS  Google Scholar 

  • Richaud C, Margarita D, Baranton G, Saint Girons I (1990) Cloning of genes required for amino acid biosynthesis from Leptospira interrogans serovar icterohaemorrhagiae. J Gen Microbiol 136:651–656

    Article  PubMed  CAS  Google Scholar 

  • Ristow P, Bourhy P, da Cruz McBride FW, Figueira CP, Huerre M, Ave P, Girons IS, Ko AI, Picardeau M (2007) The OmpA-like protein Loa22 is essential for leptospiral virulence. PLoS Pathog 3:e97

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Saint Girons I, Margarita D, Amouriaux P, Baranton G (1990) First isolation of bacteriophages for a spirochaete: potential genetic tools for Leptospira. Res Microbiol 141:1131–1138

    Article  PubMed  CAS  Google Scholar 

  • Saint Girons I, Bourhy P, Ottone C, Picardeau M, Yelton D, Hendrix RW, Glaser P, Charon N (2000) The LE1 bacteriophage replicates as a plasmid within Leptospira biflexa: construction of an L. biflexa–Escherichia coli shuttle vector. J Bacteriol 182:5700–5705

    Article  CAS  Google Scholar 

  • Satou K, Shimoji M, Tamotsu H, Juan A, Ashimine N, Shinzato M, Toma C, Nohara T, Shiroma A, Nakano K, Teruya K, Terabayashi Y, Ohki S, Koizumi N, Okano S, Suzuki T, Hirano T (2015) Complete genome sequences of low-passage virulent and high-passage avirulent variants of pathogenic Leptospira interrogans serovar manilae strain UP-MMC-NIID, originally isolated from a patient with severe leptospirosis, determined using PacBio single-molecule real-time technology. Genome Announc 3:e00815–e00882

    Article  Google Scholar 

  • Setubal JC, Reis MG, Matsunaga J, Haake DA (2006) Lipoprotein computational prediction in spirochaetal genomes. Microbiology 152:113–121

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Schuller S, Sergeant K, Renaut J, Callanan JJ, Scaife C, Nally JE (2015) Comparative proteomic analysis of lung tissue from guinea pigs with leptospiral pulmonary haemorrhage syndrome (LPHS) reveals a decrease in abundance of host proteins involved in cytoskeletal and cellular organization. J Proteomics 122:55–72

    Google Scholar 

  • Slamti L, de Pedro MA, Guichet E, Picardeau M (2011) Deciphering morphological determinants of the helix-shaped Leptospira. J Bacteriol 193:6266–6275

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Srikram A, Zhang K, Bartpho T, Lo M, Hoke DE, Sermswan RW, Adler B, Murray GL (2011) Cross-protective immunity against leptospirosis elicited by a live, attenuated lipopolysaccharide mutant. J Infect Dis 203:870–879

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Stamm LV, Parrish EA, Gherardini FC (1991) Cloning of the recA gene from a free-living leptospire and distribution of RecA-like protein among spirochetes. Appl Environ Microbiol 57:183–189

    PubMed  PubMed Central  CAS  Google Scholar 

  • Stewart PE, Carroll JA, Olano LR, Sturdevant DE, Rosa PA (2016) Multiple posttranslational modifications of Leptospira biflexa proteins as revealed by proteomic analysis. Appl Environ Microbiol 82:1183–1195

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Toma C, Murray GL, Nohara T, Mizuyama M, Koizumi N, Adler B, Suzuki T (2014) Leptospiral outer membrane protein LMB216 is involved in enhancement of phagocytic uptake by macrophages. Cell Microbiol

    Google Scholar 

  • Viratyosin W, Ingsriswang S, Pacharawongsakda E, Palittapongarnpim P (2008) Genome-wide subcellular localization of putative outer membrane and extracellular proteins in Leptospira interrogans serovar Lai genome using bioinformatics approaches. BMC Genom 9:181

    Article  CAS  Google Scholar 

  • Wang Y, Zhuang X, Zhong Y, Zhang C, Zhang Y, Zeng L, Zhu Y, He P, Dong K, Pal U, Guo X, Qin J (2015) Distribution of plasmids in distinct Leptospira pathogenic species. PLoS Negl Trop Dis 9:e0004220

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Witchell TD, Eshghi A, Nally JE, Hof R, Boulanger MJ, Wunder EAJ, Ko AI, Haake DA, Cameron CE (2014) Post-translational modification of LipL32 during Leptospira interrogans infection. PLoS Negl Trop Dis 8:e3280

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wunder EA Jr, Figueira CP, Benaroudj N, Hu BY, Tong BA, Trajtenberg F, Liu J, Reis MG, Charon NW, Buschiazzo A, Picardeau M, Ko AI (2016) A novel flagellar sheath protein, FcpA, determines filament coiling, translational motility and virulence for the Leptospira spirochete. Mol Microbiol 101:457–470

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Xu Y, Zhu Y, Wang Y, Chang YF, Zhang Y, Jiang X, Zhuang X, Zhu Y, Zhang J, Zeng L, Yang M, Li S, Wang S, Ye Q, Xin X, Zhao G, Zheng H, Guo X, Wang J (2016) Whole genome sequencing revealed host adaptation-focused genomic plasticity of pathogenic Leptospira. Sci Rep 6:20020

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Xue F, Dong H, Wu J, Wu Z, Hu W, Sun A, Troxell B, Yang XF, Yan J (2010) Transcriptional responses of Leptospira interrogans to host innate immunity: significant changes in metabolism, oxygen tolerance, and outer membrane. PLoS Negl Trop Dis 7:e2477

    Article  CAS  Google Scholar 

  • Yang HL, Zhu YZ, Qin JH, He P, Jiang XC, Zhao GP, Guo XK (2006) In silico and microarray-based genomic approaches to identifying potential vaccine candidates against Leptospira interrogans. BMC Genom 7:293

    Article  CAS  Google Scholar 

  • Yelton DB, Cohen RA (1986) Analysis of cloned DNA from Leptospira biflexa serovar patoc which complements a deletion of the Escherichia coli trpE gene. J Bacteriol 165:41–46

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhang R, Lin Y (2009) DEG 5.0, a database of essential genes in both prokaryotes and eukaryotes. Nucleic Acids Res 37:D455–D458

    Article  PubMed  CAS  Google Scholar 

  • Zhang L, Zhang C, Ojcius DM, Sun D, Zhao J, Lin X, Li L, Li L, Yan J (2012) The mammalian cell entry (Mce) protein of pathogenic Leptospira species is responsible for RGD motif-dependent infection of cells and animals. Mol Microbiol 83:1006–1023

    Article  PubMed  CAS  Google Scholar 

  • Zhang K, Murray GL, Seemann T, Srikram A, Bartpho T, Sermswan RW, Adler B, Hoke DE (2013) Leptospiral LruA is required for virulence and modulates an interaction with mammalian Apolipoprotein A-I. Infect Imm 81:3872–3879

    Article  CAS  Google Scholar 

  • Zhao JF, Chen HH, Ojcius DM, Zhao X, Sun D, Ge YM, Zheng LL, Lin X, Li LJ, Yan J (2013) Identification of Leptospira interrogans phospholipase C as a novel virulence factor responsible for intracellular free calcium ion elevation during macrophage death. PLoS ONE 8:e75652

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhong Y, Chang X, Cao XJ, Zhang Y, Zheng H, Zhu YZ, Cai C, Cui Z, Zhang Y, Li YY, Jiang XG, Zhao GP, Wang S, Li Y, Zeng R, Li X, Guo XK (2011) Comparative proteogenomic analysis of the Leptospira interrogans virulence-attenuated strain IPAV against the pathogenic strain 56601. Cell Res 21:1210–1229

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhu WN, Huang LL, Zeng LB, Zhuang XR, Chen CY, Wang YZ, Qin JH, Zhu YZ, Guo XK (2014) Isolation and characterization of two novel plasmids from pathogenic Leptospira interrogans serogroup canicola serovar canicola strain gui44. PLoS Negl Trop Dis 8:e3103

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhu W, Wang J, Zhu Y, Tang B, Zhang Y, He P, Zhang Y, Liu B, Guo X, Zhao G, Qin J (2015) Identification of three extra-chromosomal replicons in Leptospira pathogenic strain and development of new shuttle vectors. BMC Genom 16:90

    Article  CAS  Google Scholar 

  • Zhukova A, Fernandes LV, Hugon P, Pappas C, Sismeiro O, Coppée JY, Becavin C, Malabat C, Eshghi A, Zhang JJ, Yang FX, Picardeau M (2017) Genome-wide transcriptional start site mapping and sRNA identification in the pathogen Leptospira interrogans. Front Cel Infect Microbiol 7:10

    Google Scholar 

  • Zuerner RL (1991) Physical mapping of chromosomal and plasmid DNA comprising the genome of Leptospira interrogans. Nucleic Acids Res 19:4857–4860

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Picardeau, M. (2017). Toolbox of Molecular Techniques for Studying Leptospira Spp.. In: Adler, B. (eds) Spirochete Biology: The Post Genomic Era. Current Topics in Microbiology and Immunology, vol 415. Springer, Cham. https://doi.org/10.1007/82_2017_45

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