BmpA, BmpB, BmpC, and BmpD are homologous lipoproteins of unknown functions,

BmpA, BmpB, BmpC, and BmpD are homologous lipoproteins of unknown functions, encoded by the genes of paralogous chromosomal gene family 36. genome (6) and include (1, 17, 19), (21), NVP-BSK805 manufacture the 2 2.9 gene family (16), and homologue (24). While DNA of any of these gene families could be a substrate NVP-BSK805 manufacture for stochastic genetic rearrangement to yield variation in gene expression and/or antigenic variation (14), this phenomenon has not yet been demonstrated NVP-BSK805 manufacture to occur in infections. In B31, genes (paralogous gene family 36) are located in tandem in the chromosome in the order in a region extending from nucleotides 391932 to 396563 (6). They are also present in the chromosomes of other strains (1, 17, 19). In B31, DNA sequence homologies among genes range from 56 to 64%. DNA sequence analysis has suggested that is preceded by two promoters (1), that and are preceded by individual promoters (17, 19), and that is preceded by no promoter (19). The putative promoter is located within the coding sequence (1, 19). Although the functions of the proteins encoded by NVP-BSK805 manufacture the genes are unknown, organisms in culture synthesize mRNAs of all four genes (17; E. Dobrikova, V. Gorbacheva, and F. C. Cabello, unpublished data) and antibodies to BmpA, BmpC, and BmpD proteins are present in infected NVP-BSK805 manufacture hosts (1, 2, 17, 19). These data suggest that the functions of these proteins may be necessary for in vitro and in vivo growth and that at least three members of this family may have a role in virulence (4). Very few genes of that are involved in virulence have been identified as a result of obtaining the complete sequence of this organism (6). Analysis of a chromosomal region whose genes code for uncovered, putatively in vivo-induced and clearly immunogenic lipoproteins may therefore be relevant to virulence. The presence of Bmp proteins on the surface of genes are present in the genomes of all isolates of sensu lato, but at least and have been identified in and (1, 17, 19). To provide a basis for understanding the role that chromosomally encoded Bmp proteins might play in the biology of and in the pathogenesis of Lyme disease, and to evaluate the usefulness of Bmp proteins as reagents for diagnosis of Lyme disease produced KRT7 by different strains (1, 18), the structures of the regions in several species were analyzed using DNA hybridization, PCR amplification, and DNA sequencing. There were no differences in the relative order of the genes in these species, but variations in DNA sequence were significantly more common in intergenic regions than in coding regions. Bacterial strains and culture. B31 (ATCC 35210) and 297 (20); 10 sensu stricto strains recently isolated from skin biopsies and blood samples from patients with Lyme disease and passaged only once (10); G25 and N34 (from R. Marconi); Ip3 (9), ACA1 (3), VS461 (9), and VS486 (from J. Benach); 25015 (formerly sensu lato group DN127) (22); 21038 (from R. Marconi); H014 (13); and (from R. Johnson) were grown at 32 to 34C in BSK-H medium supplemented with 7% rabbit serum (Sigma Chemical Co., St. Louis, Mo.) (7). Strains were cloned by two rounds of limiting dilution in BSK-H medium or by subsurface agarose colony isolation (5). Cell concentration was determined by counting cells stained with acridine orange under fluorescence microscopy (23) or by counting viable cells on agarose plates (5). Comparable results were obtained by both techniques. Southern hybridization. Total DNA from each strain was purified from a mid-log-phase culture (8) and digested overnight with gene. Templates for these reactions were pUC19-based plasmids made up of different DNA segments of the region of 297 (2). A DNA probe targeting the gene for use as a control was obtained by PCR amplification by using total DNA purified from 297 as a template and appropriate primers (5-CTAGTGGGTACAGAATTAATCGAGC-3 and 5-GCCTGCGCAATCATTGCCATTGC-3) (11). DNA probes were purified, labeled with digoxigenin-11-dUTP by the random primer method according to the instructions of the manufacturer (Boehringer Mannheim, Indianapolis, Ind.), hybridized to DNA blots at 65C, and washed under high-stringency conditions in 0.5 SSC (1 SSC is 0.15 M NaCl plus 0.015 M sodium citrate).

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