Transcriptional analyses and mapping of the ospC gene in Lyme disease spirochetes

Marconi, Richard T.; Samuels, D. Scott; Garon, Claude F.

doi:10.1128/jb.175.4.926-932.1993

Cited by 166 publications

(198 citation statements)

References 28 publications

(17 reference statements)

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“…This plasmid might have been cp32-1, which contains the erpAB locus and was cut by BamHI to produce fragments of approximately 22 and 10 kb. Samuels and coworkers (28,40) have also observed a circular plasmid in B. burgdorferi B31 that migrated in twodimensional gels with a mobility slightly slower than that of the 26-kb ospC-guaA plasmid, as was found for the 32-kb plasmids described in this work. In addition, Porcella et al (36) have cloned several related copies of a sequence from B. burgdorferi 297 that hybridize to an approximately 30-kb circular plasmid, suggesting that this isolate may also contain multiple circular plasmids of this size.…”

Section: Resultsmentioning

confidence: 50%

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A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

1996

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We have identified four loci in Borrelia burgdorferi B31 that contain open reading frames capable of encoding six proteins that are related to the antigenic proteins OspE and OspF. We have designated these proteins Erp, for OspEF-related protein, and named their respective genes erp. The erpA and erpB genes are linked, as are erpC and erpD, and the pairs probably constitute two operons. The erpG and erpH genes appear to be monocistronic. The ErpA and ErpC proteins are expressed by B. burgdorferi B31 in culture and are recognized by a polyclonal antiserum raised against the OspE protein of B. burgdorferi N40. The four erp loci are each located on different 32-kb circular plasmids that contain additional DNA sequences that are homologous to each other and to an 8.3-kb circular plasmid of B. burgdorferi sensu lato Ip21. All four 32-kb plasmids can be maintained within a single bacterium, which may provide a model for the study of plasmid replication and segregation in B. burgdorferi.Borrelia burgdorferi is a member of the order Spirochaetales, the spirochetes, an order that is phylogenetically and morphologically distinct from such well-characterized bacteria as Escherichia coli and Bacillus subtilis (8,24,34,35,53). B. burgdorferi bacteria naturally contain a wide variety of plasmids (4,13,25,40,42,45,47,54), yet nothing is known about the mechanisms by which plasmids are maintained in these bacteria. Several observations suggest that the small DNA species of B. burgdorferi and other spirochetes may be fundamentally different from the plasmids of other bacteria, the most significant being that there are linear plasmids in B. burgdorferi (4,5,7,13,22,23,42). Also, two B. burgdorferi genes involved in purine biosynthesis are located on a circular plasmid (29), which appears to be inconsistent with the classical definition of a plasmid as a nonessential extrachromosomal element. Other plasmids of B. burgdorferi appear to be capable of undergoing recombination to form multimeric plasmids (25, 31), and there have been reports of DNA sequences located on several different plasmids within the same bacterium (46,47,(54)(55)(56).B. burgdorferi is the causative agent of Lyme disease, an increasingly common ailment of humans and several other mammals (15,48). Infection of a mammal by B. burgdorferi is generally accompanied by the production of antibodies directed against a limited number of bacterial antigens (17,19,41,52), several of which have been identified as surface-exposed lipoproteins. At least three of these lipoproteins, OspC, OspE, and OspF, are differentially expressed by B. burgdorferi, being produced in greater quantities by cultured bacteria that are shifted from 23 to 32ЊC than by those maintained at the lower temperature (43, 49). For OspC, at least, this appears to be related to induction of specific protein synthesis by the spirochete in ticks following feeding upon a warm-blooded animal (43). Temperature-dependent differential expression of bacterial proteins that are involved in host infection has been o...

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Section: Resultsmentioning

confidence: 50%

“…The filter was stripped and rehybridized with a probe specific for guaA, which is located on the same 26-kb circular plasmid as ospC (28,29,39). This probe hybridized to a linear molecule of approximately 26 kb and to two circular DNAs (Fig.…”

Section: Resultsmentioning

confidence: 99%

A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

1996

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“…In light of the notion that needle exposure by no means reflects the situation of tick infection [23,25,54] and the highly improved real-time PCR technique to quantify B. burgdorferi in situ [24,27,43,46,48,62,63,64], we have now performed a detailed quantitative analysis of the spirochete burden and population dynamics in different tissues of mice following natural (tick) infection by B. burgdorferi s.s. (ZS7) for a period of 4 months postinfection (p.i.). For this purpose we have selected one cp26-encoded gene, ospC [40], and four previously described lp54-encoded genes, i.e. ospA, zs7.a36, zs7.a66, and zs7.a68 [20,61]-the respective antigens of which are suggested to be either putative vaccine candidates [61,66] and/or relevant for complement resistence [32]-and three strains of mice with distinct immune status and susceptibility to B. burgdorferi s.s.-induced disease/ infection [50,53].…”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis of Borrelia burgdorferi gene expression in naturally (tick) infected mouse strains

Lederer

Brenner

Stehlé

et al. 2004

Med Microbiol Immunol

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Adaptation of Borrelia burgdorferi in the vector and vertebrate host is mediated by mechanisms that regulate differential expression of outer surface lipoproteins (Osps). In this study, real time PCR was applied to quantify tissue-specific expression of four linear plasmid (lp54)-encoded (ospA, zs7.a36, zs7.a66 zs7.a68) and one circular plasmid (cp26)-encoded (ospC) gene from B. burgdorferi sensu stricto, in a natural setting of tick-infected immunodeficient (C.B-17 SCID) and immunocompetent (BALB/c and AKR/OlaHsd) mice for up to 120 days post-infection (p.i.). Early during infection (day 30 p.i.) high numbers of spirochetes were found in the heart and joint, but not the ear and spleen tissues of disease-susceptible SCID mice. In diseasesusceptible AKR mice spirochetes colonized the ear and joint tissues, but were undetectable in tissues of diseaseresistant BALB/c mice. Later in infection (day 120 p.i.), spirochetes had expanded ($1,000-fold) in all SCID tissues tested but were undetectable in AKR and BALB/ c mice. Of the five genes analyzed, only zs7.a36 transcripts were detected in various tissues of all infected mouse strains, though at differing levels, whereas ospC transcripts were only found in tissue specimens of SCID mice. Furthermore, gene expression of ospC and zs7.a36 appears to be differentially regulated in distinct organs of individual mice. In contrast, transcripts for ospA, zs7.a66, and zs7.a68 were not detected in any of the mouse strains, independent of their immune status and/ or the severity of their infection/inflammatory responses. Late during infection (day 120 p.i.), transcription of zs7.a36 and ospC was down-regulated in the tissues of SCID mice despite expansion of spirochetes. This type of quantitative analysis may be helpful to further disclose principles of pathogenesis of Lyme borreliosis and to design strategies for its therapeutic treatment.

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“…The outer surface protein C gene (ospC ) of B. burgdorferi sensu lato is located on a 27-kb circular plasmid [25] and is highly heterogeneous with the nucleotide sequences differing among isolates of different species of Borrelia [26,27]. Strain diversity as well as the genetic heterogeneity can be distinguished among different Borrelia isolates by their distinct RFLP types [28].…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis of the outer surface protein C gene of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan

Shih

Chao

2002

Journal of Medical Microbiology

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The outer surface protein C gene (ospC) of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) was analysed for the first time in Taiwan. The genetic identities of these Taiwan isolates (TWKM1-7) were determined by restriction fragment length polymorphism (RFLP) analysis and sequence similarities of the PCR-amplified ospC gene amplicons. After cleavage by nuclease Dra I, differential fragment patterns of PCRamplified ospC DNA in relation to different genospecies of Lyme disease spirochaetes were observed and all of these Taiwan isolates were genetically aff iliated to the genospecies of B. burgdorferi sensu stricto. The phylogenetic analysis on the sequence similarity of these Taiwan isolates revealed a highly homogeneous genotype, ranging from 99.3% to 100%, within the genospecies of B. burgdorferi sensu stricto and was distinguished from other genospecies of Borrelia isolates. The sequence similarity analysis also revealed the high sequence variability of the ospC gene among Borrelia strains that belong to the same genospecies but were isolated from different biological and geographical sources. Thus, these results provide the first investigation on the genetic identity of the ospC gene of these Taiwan isolates and show that these Taiwan isolates were closely related genetically to the genospecies of B. burgdorferi sensu stricto.

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Transcriptional analyses and mapping of the ospC gene in Lyme disease spirochetes

Cited by 166 publications

References 28 publications

A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

Quantitative analysis of Borrelia burgdorferi gene expression in naturally (tick) infected mouse strains

Genetic analysis of the outer surface protein C gene of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan

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