13. Mitchison M, Bulach DM, Vinh T,
Rajakumar K, Faine S, Adler B (1997) Identification and characterization of the dTDPrhamnose biosynthesis and transfer genes of
the lipopolysaccharide-related rfb locus in Leptospira interrogans serovar Copenhageni. J
Bacteriol 179(4):1262–1267. https://doi.
org/10.1128/jb.179.4.1262-1267.1997
14. Girons IS, 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
15. Bauby H, Saint Girons I, Picardeau M (2003)
Construction and complementation of the first
auxotrophic mutant in the spirochaete Leptospira meyeri. Microbiology 149(Pt 3):689–693.
https://doi.org/10.1099/mic.0.26065-0
16. Bourhy P, Louvel H, Saint Girons I, Picardeau
M (2005) Random insertional mutagenesis of
Leptospira interrogans, the agent of leptospirosis, using a mariner transposon. J Bacteriol
187:3255–3258. 187/9/3255 [pii]
17. Picardeau M (2008) Conjugative transfer
between Escherichia coli and Leptospira spp. as
a new genetic tool. Appl Environ Microbiol
74:319–322. AEM.02172-07 [pii]
18. Murray GL, Morel V, Cerqueira GM, Croda J,
Srikram A, Henry R, Ko AI, Dellagostin OA,
Bulach DM, Sermswan RW, Adler B, Picardeau
M (2009) Genome-wide transposon mutagenesis in pathogenic Leptospira species. Infect
Immun 77:810–816. https://doi.org/10.
1128/IAI.01293-08
19. Poggi D, De Giuseppe PO, Picardeau M
(2010) Antibiotic resistance markers for
genetic manipulations of Leptospira spp. Appl
Environ
Microbiol
76(14):4882–4885.
https://doi.org/10.1128/AEM.00775-10
20. Pappas CJ, Benaroudj N, Picardeau M (2015)
A replicative plasmid vector allows efficient
complementation of pathogenic Leptospira
strains.
Appl
Environ
Microbiol
81:3176–3181.
https://doi.org/10.1128/
AEM.00173-15
21. 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 extrachromosomal replicons in Leptospira pathogenic strain and development of new shuttle
vectors. BMC Genomics 16:90. https://doi.
org/10.1186/s12864-015-1321-y
22. 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(1):189–199. https://doi.
org/10.1046/j.1365-2958.2001.02374.x
23. Kameni APT, Couture-Tosi E, Saint-Girons I,
Picardeau M (2002) Inactivation of the spirochete recA gene results in a mutant with low
viability and irregular nucleoid morphology. J
Bacteriol 184(2):452–458. https://doi.org/
10.1128/JB.184.2.452-458.2002
24. Gue ´gan R, Camadro JM, Saint Girons I, Picardeau M (2003) Leptospira spp. possess a complete haem biosynthetic pathway and are able
to use exogenous haem sources. Mol Microbiol
49(3):745–754. https://doi.org/10.1046/j.
1365-2958.2003.03589.x
25. Picardeau M, Bauby H, Saint Girons I (2003)
Genetic evidence for the existence of two pathways for the biosynthesis of methionine in the
Leptospira spp. FEMS Microbiol Lett 225
(2):257–262.
https://doi.org/10.1016/
S0378-1097(03)00529-9
26. Croda J, Figueira CP, Wunder EA, Santos CS,
Reis MG, Ko AI, Picardeau M (2008) Targeted
mutagenesis in pathogenic Leptospira species:
disruption of the LigB gene does not affect
virulence in animal models of leptospirosis.
Infect Immun 76(12):5826–5833. https://
doi.org/10.1128/IAI.00989-08
27. Craig NL (1997) Target site selection in transposition. Annu Rev Biochem 66:437–474.
https://doi.org/10.1146/annurev.biochem.
66.1.437
28. Rubin EJ, Akerley BJ, Novik VN, Lampe DJ,
Husson RN, Mekalanos JJ (1999) In vivo
transposition of mariner-based elements in
enteric bacteria and mycobacteria. Proc Natl
Acad Sci U S A 96(4):1645–1650
29. Zhang JK, Pritchett MA, Lampe DJ, Robertson HM, Metcalf WW (2000) In vivo transposon mutagenesis of the methanogenic archaeon
Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1. Proc Natl Acad Sci U
S A 97(17):9665–9670. https://doi.org/10.
1073/pnas.160272597
30. Ashour J, Hondalus MK (2003) Phenotypic
mutants of the intracellular actinomycete Rhodococcus equi created by in vivo Himar1 transposon
mutagenesis.
J
Bacteriol
185:2644–2652. https://doi.org/10.1128/
JB.185.8.2644-2652.2003
31. Lamichhane G, Zignol M, Blades NJ, Geiman
DE, Dougherty A, Grosset J, Broman KW,
Bishai WR (2003) A postgenomic method for
predicting essential genes at subsaturation
levels of mutagenesis: application to Mycobacterium tuberculosis. Proc Natl Acad Sci U S A
100(12):7213–7218.
https://doi.org/10.
1073/pnas.1231432100
Random Transposon Mutant Library
95
Rajakumar K, Faine S, Adler B (1997) Identification and characterization of the dTDPrhamnose biosynthesis and transfer genes of
the lipopolysaccharide-related rfb locus in Leptospira interrogans serovar Copenhageni. J
Bacteriol 179(4):1262–1267. https://doi.
org/10.1128/jb.179.4.1262-1267.1997
14. Girons IS, 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
15. Bauby H, Saint Girons I, Picardeau M (2003)
Construction and complementation of the first
auxotrophic mutant in the spirochaete Leptospira meyeri. Microbiology 149(Pt 3):689–693.
https://doi.org/10.1099/mic.0.26065-0
16. Bourhy P, Louvel H, Saint Girons I, Picardeau
M (2005) Random insertional mutagenesis of
Leptospira interrogans, the agent of leptospirosis, using a mariner transposon. J Bacteriol
187:3255–3258. 187/9/3255 [pii]
17. Picardeau M (2008) Conjugative transfer
between Escherichia coli and Leptospira spp. as
a new genetic tool. Appl Environ Microbiol
74:319–322. AEM.02172-07 [pii]
18. Murray GL, Morel V, Cerqueira GM, Croda J,
Srikram A, Henry R, Ko AI, Dellagostin OA,
Bulach DM, Sermswan RW, Adler B, Picardeau
M (2009) Genome-wide transposon mutagenesis in pathogenic Leptospira species. Infect
Immun 77:810–816. https://doi.org/10.
1128/IAI.01293-08
19. Poggi D, De Giuseppe PO, Picardeau M
(2010) Antibiotic resistance markers for
genetic manipulations of Leptospira spp. Appl
Environ
Microbiol
76(14):4882–4885.
https://doi.org/10.1128/AEM.00775-10
20. Pappas CJ, Benaroudj N, Picardeau M (2015)
A replicative plasmid vector allows efficient
complementation of pathogenic Leptospira
strains.
Appl
Environ
Microbiol
81:3176–3181.
https://doi.org/10.1128/
AEM.00173-15
21. 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 extrachromosomal replicons in Leptospira pathogenic strain and development of new shuttle
vectors. BMC Genomics 16:90. https://doi.
org/10.1186/s12864-015-1321-y
22. 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(1):189–199. https://doi.
org/10.1046/j.1365-2958.2001.02374.x
23. Kameni APT, Couture-Tosi E, Saint-Girons I,
Picardeau M (2002) Inactivation of the spirochete recA gene results in a mutant with low
viability and irregular nucleoid morphology. J
Bacteriol 184(2):452–458. https://doi.org/
10.1128/JB.184.2.452-458.2002
24. Gue ´gan R, Camadro JM, Saint Girons I, Picardeau M (2003) Leptospira spp. possess a complete haem biosynthetic pathway and are able
to use exogenous haem sources. Mol Microbiol
49(3):745–754. https://doi.org/10.1046/j.
1365-2958.2003.03589.x
25. Picardeau M, Bauby H, Saint Girons I (2003)
Genetic evidence for the existence of two pathways for the biosynthesis of methionine in the
Leptospira spp. FEMS Microbiol Lett 225
(2):257–262.
https://doi.org/10.1016/
S0378-1097(03)00529-9
26. Croda J, Figueira CP, Wunder EA, Santos CS,
Reis MG, Ko AI, Picardeau M (2008) Targeted
mutagenesis in pathogenic Leptospira species:
disruption of the LigB gene does not affect
virulence in animal models of leptospirosis.
Infect Immun 76(12):5826–5833. https://
doi.org/10.1128/IAI.00989-08
27. Craig NL (1997) Target site selection in transposition. Annu Rev Biochem 66:437–474.
https://doi.org/10.1146/annurev.biochem.
66.1.437
28. Rubin EJ, Akerley BJ, Novik VN, Lampe DJ,
Husson RN, Mekalanos JJ (1999) In vivo
transposition of mariner-based elements in
enteric bacteria and mycobacteria. Proc Natl
Acad Sci U S A 96(4):1645–1650
29. Zhang JK, Pritchett MA, Lampe DJ, Robertson HM, Metcalf WW (2000) In vivo transposon mutagenesis of the methanogenic archaeon
Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1. Proc Natl Acad Sci U
S A 97(17):9665–9670. https://doi.org/10.
1073/pnas.160272597
30. Ashour J, Hondalus MK (2003) Phenotypic
mutants of the intracellular actinomycete Rhodococcus equi created by in vivo Himar1 transposon
mutagenesis.
J
Bacteriol
185:2644–2652. https://doi.org/10.1128/
JB.185.8.2644-2652.2003
31. Lamichhane G, Zignol M, Blades NJ, Geiman
DE, Dougherty A, Grosset J, Broman KW,
Bishai WR (2003) A postgenomic method for
predicting essential genes at subsaturation
levels of mutagenesis: application to Mycobacterium tuberculosis. Proc Natl Acad Sci U S A
100(12):7213–7218.
https://doi.org/10.
1073/pnas.1231432100
Random Transposon Mutant Library
95