Mol Microbiol 1990,4(11):1911–1919.CrossRefPubMed 26. Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: a laboratory manual. 2 Edition Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press 1989. 27. Timm J, Lim EM, Gicquel B:Escherichia coli -mycobacteria RO4929097 nmr shuttle vectors for operon and gene fusions to lacZ : the pJEM series. J Bacteriol 1994,176(21):6749–6753.PubMed 28. Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR: Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 1989,77(1):51–59.CrossRefPubMed
29. Pelicic V, Jackson M, Reyrat JM, Jacobs WR Jr, Gicquel B, Guilhot C: Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis. Proc Natl Acad Sci USA 1997,94(20):10955–10960.CrossRefPubMed 30. Hanahan D, Jessee J, Bloom FR: Plasmid transformation of Escherichia coli and other bacteria. Methods Enzymol 1991, 204:63–113.CrossRefPubMed Authors’ contributions SG contributed to design of the study, participated in growth experiments, phosphate
transport and reporter gene assays and drafted the manuscript. NE carried out the molecular work and participated in all other experimental aspects. GMC contributed to design of the study, participated in phosphate transport assays and helped to draft the manuscript. All authors read and approved the final manuscript.”
“Background The genus Leptospira selleck compound is composed of both saprophytic and pathogenic species . Pathogenic Leptospira spp., such as L. interrogans, L. borgpetersenii,
L. weilii and L. kirschner, are the causative agents of leptospirosis, a serious world-wide disease in humans and animals [2, 3]. The disease in humans occurs mostly after contact, often through skin wounds, with soil or water contaminated Adenosine by urine of infected animals. Its severity varies from mild to rapidly fatal. Severe symptoms are characterized by visible jaundice involving hepatic injury, acute renal failure, carditis and hemorrhage, and case fatality varies from a few percent to 25% [3–6]. However, the mechanisms of disease caused by pathogenic Leptospira spp. remain largely unknown. Both pathogenic and saprophytic leptospires express two endoflagella (periplasmic flagella). One of the endoflagella is attached at one end of the cell and is located between the protoplasmic cylinder and the outer membrane sheath [7–9]. The endoflagella, rotating within the periplasmic space, are responsible for spirochete motility. In pathogenic Leptospira species, this motility is considered to contribute to invasion into hosts and diffusion within the hosts during infection [9, 10]. In previous studies, we found that pathogenic leptospires can adhere to host cells with one or two termini of the microbial bodies, while Semaxanib supplier non-pathogenic leptospiral strains lacked this ability [11, 12].