b31  (ATCC)


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    Structured Review

    ATCC b31
    In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from <t>B31-A-derivative</t> strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.
    B31, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/b31/product/ATCC
    Average 92 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    b31 - by Bioz Stars, 2022-09
    92/100 stars

    Images

    1) Product Images from "Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi"

    Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2007.05969.x

    In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from B31-A-derivative strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.
    Figure Legend Snippet: In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from B31-A-derivative strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.

    Techniques Used: In Vitro, Standard Deviation, Two Tailed Test

    Expression and cellular localization of the BBB26-FLAG and BBB27-FLAG proteins. A. Proteins lysates from E. coli (Ec) or B. burgdorferi B31-A34 (Bb) harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were separated by SDS-PAGE and analysed by immunoblot with anti-FLAG antibodies. The mobilities of size standards (molecular weights in kDa) are indicated to the left of the figure. B. Protein lysates from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were harvested and separated into soluble and membrane fractions by ultracentrifugation. Protein fractions from equivalent numbers of spirochetes were subjected to SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), OppAIV (inner membrane), OspB (outer membrane) and SodA (cytoplasmic) antisera. Representative results for the localization of the OppAIV, OspB and SodA proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown. L, total cell lysate; S, soluble protein fraction; M, membrane protein fraction. C. Equal numbers of whole or 0.1% SDS-treated (+0.1% SDS) cells of B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were incubated with different concentrations (μg ml −1 ) of proteinase K (PK). Lysates of PK-treated bacteria were separated by SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), flagellin (FlaB, periplasmic marker), OppAIV (inner membrane marker) and OspB (surface exposed, outer membrane marker) antisera. Representative results for the proteinase K sensitivity of the FlaB, OppAIV and OspB proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown.
    Figure Legend Snippet: Expression and cellular localization of the BBB26-FLAG and BBB27-FLAG proteins. A. Proteins lysates from E. coli (Ec) or B. burgdorferi B31-A34 (Bb) harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were separated by SDS-PAGE and analysed by immunoblot with anti-FLAG antibodies. The mobilities of size standards (molecular weights in kDa) are indicated to the left of the figure. B. Protein lysates from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were harvested and separated into soluble and membrane fractions by ultracentrifugation. Protein fractions from equivalent numbers of spirochetes were subjected to SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), OppAIV (inner membrane), OspB (outer membrane) and SodA (cytoplasmic) antisera. Representative results for the localization of the OppAIV, OspB and SodA proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown. L, total cell lysate; S, soluble protein fraction; M, membrane protein fraction. C. Equal numbers of whole or 0.1% SDS-treated (+0.1% SDS) cells of B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were incubated with different concentrations (μg ml −1 ) of proteinase K (PK). Lysates of PK-treated bacteria were separated by SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), flagellin (FlaB, periplasmic marker), OppAIV (inner membrane marker) and OspB (surface exposed, outer membrane marker) antisera. Representative results for the proteinase K sensitivity of the FlaB, OppAIV and OspB proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown.

    Techniques Used: Expressing, SDS Page, Incubation, Marker

    A. PCR analysis of genomic DNA from B. burgdorferi clones transformed with gene inactivation constructs targeting genes in the bbb26–27 region. Template DNAs from transformants are identified below the lanes and PCR amplification targets above the lanes. Template DNA from B31-A illustrates the PCR products from the wild-type alleles of bbb26–27 , bbb26 and bbb27 (lanes 2–5, 15 and 19), whereas template DNAs from the gene inactivation constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ) depict the PCR profiles of the mutated alleles (lanes 6–9, 16 and 20). The PCR products resulting from the clones transformed with the allelic exchange inactivation plasmids are illustrated in lanes 10–13, 17 and 21. The 1 kbp-plus size standards (Invitrogen) were run in lanes 1, 14 and 18 and sizes (base pairs) are indicated to the left of the panel. B. Graphical representation of the bbb26–27 region on cp26 (B31-A) and the cloned pieces of DNA used for the allelic exchange constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ). The 1168 bp region of cp26 between nucleotides 21923 and 23091 was replaced with the 1146 bp flaB p – aadA resistance cassette to create XL-BBB26–27Δ for disruption of both bbb26 and bbb27 . The 742 bp region of cp26 between nucleotides 21923 and 22579 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB26Δ for disruption of bbb26 . The 438 bp region of cp26 between nucleotides 22653 and 23091 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB27Δ for disruption of bbb27 . Locations of the primers used for analysis in (A) are indicated and the sequences are listed in Table S1 .
    Figure Legend Snippet: A. PCR analysis of genomic DNA from B. burgdorferi clones transformed with gene inactivation constructs targeting genes in the bbb26–27 region. Template DNAs from transformants are identified below the lanes and PCR amplification targets above the lanes. Template DNA from B31-A illustrates the PCR products from the wild-type alleles of bbb26–27 , bbb26 and bbb27 (lanes 2–5, 15 and 19), whereas template DNAs from the gene inactivation constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ) depict the PCR profiles of the mutated alleles (lanes 6–9, 16 and 20). The PCR products resulting from the clones transformed with the allelic exchange inactivation plasmids are illustrated in lanes 10–13, 17 and 21. The 1 kbp-plus size standards (Invitrogen) were run in lanes 1, 14 and 18 and sizes (base pairs) are indicated to the left of the panel. B. Graphical representation of the bbb26–27 region on cp26 (B31-A) and the cloned pieces of DNA used for the allelic exchange constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ). The 1168 bp region of cp26 between nucleotides 21923 and 23091 was replaced with the 1146 bp flaB p – aadA resistance cassette to create XL-BBB26–27Δ for disruption of both bbb26 and bbb27 . The 742 bp region of cp26 between nucleotides 21923 and 22579 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB26Δ for disruption of bbb26 . The 438 bp region of cp26 between nucleotides 22653 and 23091 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB27Δ for disruption of bbb27 . Locations of the primers used for analysis in (A) are indicated and the sequences are listed in Table S1 .

    Techniques Used: Polymerase Chain Reaction, Clone Assay, Transformation Assay, Construct, Amplification

    2) Product Images from "RNA-Seq-based analysis of changes in Borrelia burgdorferi gene expression linked to pathogenicity"

    Article Title: RNA-Seq-based analysis of changes in Borrelia burgdorferi gene expression linked to pathogenicity

    Journal: Parasites & Vectors

    doi: 10.1186/s13071-014-0623-2

    Functional categories of genes upregulated in B. burgdorferi B31.
    Figure Legend Snippet: Functional categories of genes upregulated in B. burgdorferi B31.

    Techniques Used: Functional Assay

    Expression profiles of genes encoding Borrelia membrane proteins detected by qRT-PCR. Each bar represents the fold change of gene expression in B. garinii SZ vs. B. burgdorferi B31. Expression levels were nomalized to that of fla B, and levels in B. burgdorferi B31 were used to calculate fold change based on a mean of three biological replicates. Bars above and below the x-axis show genes that are up- and downregulated, respectively, in B. garinii SZ. ΔΔCt values were analyzed with the Student’s t test. *P
    Figure Legend Snippet: Expression profiles of genes encoding Borrelia membrane proteins detected by qRT-PCR. Each bar represents the fold change of gene expression in B. garinii SZ vs. B. burgdorferi B31. Expression levels were nomalized to that of fla B, and levels in B. burgdorferi B31 were used to calculate fold change based on a mean of three biological replicates. Bars above and below the x-axis show genes that are up- and downregulated, respectively, in B. garinii SZ. ΔΔCt values were analyzed with the Student’s t test. *P

    Techniques Used: Expressing, Quantitative RT-PCR

    Average log 2 -transformed reads per kilobase per million of genes differentially expressed by B. garinii SZ (y-axis) and B. burgdorferi B31 (x-axis). Red and green dots represent genes that are significantly up- and downregulated, respectively, in B. garinii SZ; gray dots represent genes that are not differentially expressed between the two species.
    Figure Legend Snippet: Average log 2 -transformed reads per kilobase per million of genes differentially expressed by B. garinii SZ (y-axis) and B. burgdorferi B31 (x-axis). Red and green dots represent genes that are significantly up- and downregulated, respectively, in B. garinii SZ; gray dots represent genes that are not differentially expressed between the two species.

    Techniques Used: Transformation Assay

    3) Product Images from "Evaluation of in-vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi"

    Article Title: Evaluation of in-vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi

    Journal: Infection and Drug Resistance

    doi: 10.2147/IDR.S19201

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values

    Techniques Used: Microscopy

    Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Effect of doxycycline, tinidazole, tigecycline, metronidazole, and amoxicillin on spirochete forms of strain B31 (top panel) and strain S297 (bottom panel). Notes: * P values calculated were
    Figure Legend Snippet: Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Effect of doxycycline, tinidazole, tigecycline, metronidazole, and amoxicillin on spirochete forms of strain B31 (top panel) and strain S297 (bottom panel). Notes: * P values calculated were

    Techniques Used: Microscopy, Staining

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to the most effective concentrations of three antibiotics measured by dark-field microscopy. Tinidazole, metronidazole, and doxycycline effect on B. burgdorferi after 3 weeks of subculturing following 72-hour treatment. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to the most effective concentrations of three antibiotics measured by dark-field microscopy. Tinidazole, metronidazole, and doxycycline effect on B. burgdorferi after 3 weeks of subculturing following 72-hour treatment. Note: * P values

    Techniques Used: Microscopy, Subculturing Assay

    Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Visualization of spirochete and round body forms of strain B31 following antibiotic treatment measured by dark field microscopy: (Ca) Control; (Cb) Doxycycline; (Cc) Tinidazole; (Cd) Metronidazole; (Ce) Tigecycline; (Cf) Amoxicillin. Note: All images taken at 40× magnification.
    Figure Legend Snippet: Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Visualization of spirochete and round body forms of strain B31 following antibiotic treatment measured by dark field microscopy: (Ca) Control; (Cb) Doxycycline; (Cc) Tinidazole; (Cd) Metronidazole; (Ce) Tigecycline; (Cf) Amoxicillin. Note: All images taken at 40× magnification.

    Techniques Used: Microscopy, Staining

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values

    Techniques Used: Microscopy

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to the most effective concentrations of three antibiotics measured by dark-field microscopy. Tinidazole, metronidazole, and doxycycline effect on B. burgdorferi after 72-hour treatment. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to the most effective concentrations of three antibiotics measured by dark-field microscopy. Tinidazole, metronidazole, and doxycycline effect on B. burgdorferi after 72-hour treatment. Note: * P values

    Techniques Used: Microscopy

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values

    Techniques Used: Microscopy

    Evaluation of biofilm-like colonies of B. burgdorferi . Quantitative analysis of biofilm-like colonies of strain B31 (top panel) and strain S297 (bottom panel) measured by crystal violet staining technique.
    Figure Legend Snippet: Evaluation of biofilm-like colonies of B. burgdorferi . Quantitative analysis of biofilm-like colonies of strain B31 (top panel) and strain S297 (bottom panel) measured by crystal violet staining technique.

    Techniques Used: Staining

    Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Effect of doxycycline, tinidazole, tigecycline, metronidazole and amoxicillin on round body forms of strain B31 (top panel) and strain S297 (bottom panel). Notes: * P values calculated were
    Figure Legend Snippet: Evaluation of live/dead spirochete and round body forms of B. burgdorferi following treatment with five antibiotics measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms). Effect of doxycycline, tinidazole, tigecycline, metronidazole and amoxicillin on round body forms of strain B31 (top panel) and strain S297 (bottom panel). Notes: * P values calculated were

    Techniques Used: Microscopy, Staining

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values

    Techniques Used: Microscopy

    Evaluation of biofilm-like colonies of B. burgdorferi . Qualitative analysis of biofilm-like colonies of strain B31 measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms): (Ba) Control; (Bb) Doxycycline; (Bc) Tinidazole; (Bd) Tigecycline; (Be) Metronidazole; (Bf) Amoxicillin. Note: All images taken at 40× magnification.
    Figure Legend Snippet: Evaluation of biofilm-like colonies of B. burgdorferi . Qualitative analysis of biofilm-like colonies of strain B31 measured by fluorescent microscopy using SYTO ® 9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms): (Ba) Control; (Bb) Doxycycline; (Bc) Tinidazole; (Bd) Tigecycline; (Be) Metronidazole; (Bf) Amoxicillin. Note: All images taken at 40× magnification.

    Techniques Used: Microscopy, Staining

    Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values
    Figure Legend Snippet: Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to different concentrations (between calculated MIC and MBC) of five antibiotics after 72-hour treatment measured by dark-field microscopy. Note: * P values

    Techniques Used: Microscopy

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  • b31  (ATCC)
    92
    ATCC b31
    In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from <t>B31-A-derivative</t> strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.
    B31, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/b31/product/ATCC
    Average 92 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    b31 - by Bioz Stars, 2022-09
    92/100 stars
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    In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from B31-A-derivative strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.

    Journal: Molecular Microbiology

    Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi

    doi: 10.1111/j.1365-2958.2007.05969.x

    Figure Lengend Snippet: In vitro growth of B. burgdorferi strains with different cp26 gene complements. The presence and absence of cp26, resT , bbb26 and bbb27 from B31-A-derivative strains are as indicated. The densities of cultures from a starting dilution of 1 × 10 5 spirochetes ml −1 were determined every 24 h using a Petroff–Hausser counting chamber. Data are presented as averages of at least three separate experiments, with triplicate cultures per strain in each experiment, and error bars represent standard deviation from the mean. Statistical analyses (Student's t -test, two tailed) were used to compare pairs of data points. The exponential phase growth of BbRB4 was significantly slower than that of all other strains, as indicated by asterisks at 24, 49 and 72 h; the P -values represent comparisons with BbRB3. All three strains lacking cp26 reached significantly lower stationary phase densities relative to B31-A, as indicated by the asterisk and P -value at 153 h.

    Article Snippet: B31 clone A (B31-A) ( ) is a non-infectious derivative of type strain B31 (ATCC 35210), which was isolated from a tick collected on Shelter Island in New York ( ).

    Techniques: In Vitro, Standard Deviation, Two Tailed Test

    Expression and cellular localization of the BBB26-FLAG and BBB27-FLAG proteins. A. Proteins lysates from E. coli (Ec) or B. burgdorferi B31-A34 (Bb) harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were separated by SDS-PAGE and analysed by immunoblot with anti-FLAG antibodies. The mobilities of size standards (molecular weights in kDa) are indicated to the left of the figure. B. Protein lysates from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were harvested and separated into soluble and membrane fractions by ultracentrifugation. Protein fractions from equivalent numbers of spirochetes were subjected to SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), OppAIV (inner membrane), OspB (outer membrane) and SodA (cytoplasmic) antisera. Representative results for the localization of the OppAIV, OspB and SodA proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown. L, total cell lysate; S, soluble protein fraction; M, membrane protein fraction. C. Equal numbers of whole or 0.1% SDS-treated (+0.1% SDS) cells of B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were incubated with different concentrations (μg ml −1 ) of proteinase K (PK). Lysates of PK-treated bacteria were separated by SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), flagellin (FlaB, periplasmic marker), OppAIV (inner membrane marker) and OspB (surface exposed, outer membrane marker) antisera. Representative results for the proteinase K sensitivity of the FlaB, OppAIV and OspB proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown.

    Journal: Molecular Microbiology

    Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi

    doi: 10.1111/j.1365-2958.2007.05969.x

    Figure Lengend Snippet: Expression and cellular localization of the BBB26-FLAG and BBB27-FLAG proteins. A. Proteins lysates from E. coli (Ec) or B. burgdorferi B31-A34 (Bb) harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were separated by SDS-PAGE and analysed by immunoblot with anti-FLAG antibodies. The mobilities of size standards (molecular weights in kDa) are indicated to the left of the figure. B. Protein lysates from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were harvested and separated into soluble and membrane fractions by ultracentrifugation. Protein fractions from equivalent numbers of spirochetes were subjected to SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), OppAIV (inner membrane), OspB (outer membrane) and SodA (cytoplasmic) antisera. Representative results for the localization of the OppAIV, OspB and SodA proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown. L, total cell lysate; S, soluble protein fraction; M, membrane protein fraction. C. Equal numbers of whole or 0.1% SDS-treated (+0.1% SDS) cells of B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG were incubated with different concentrations (μg ml −1 ) of proteinase K (PK). Lysates of PK-treated bacteria were separated by SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), flagellin (FlaB, periplasmic marker), OppAIV (inner membrane marker) and OspB (surface exposed, outer membrane marker) antisera. Representative results for the proteinase K sensitivity of the FlaB, OppAIV and OspB proteins from B. burgdorferi clone A34 harbouring either pBSV2ex bbb26 -FLAG or pBSV2ex bbb27 -FLAG are shown.

    Article Snippet: B31 clone A (B31-A) ( ) is a non-infectious derivative of type strain B31 (ATCC 35210), which was isolated from a tick collected on Shelter Island in New York ( ).

    Techniques: Expressing, SDS Page, Incubation, Marker

    A. PCR analysis of genomic DNA from B. burgdorferi clones transformed with gene inactivation constructs targeting genes in the bbb26–27 region. Template DNAs from transformants are identified below the lanes and PCR amplification targets above the lanes. Template DNA from B31-A illustrates the PCR products from the wild-type alleles of bbb26–27 , bbb26 and bbb27 (lanes 2–5, 15 and 19), whereas template DNAs from the gene inactivation constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ) depict the PCR profiles of the mutated alleles (lanes 6–9, 16 and 20). The PCR products resulting from the clones transformed with the allelic exchange inactivation plasmids are illustrated in lanes 10–13, 17 and 21. The 1 kbp-plus size standards (Invitrogen) were run in lanes 1, 14 and 18 and sizes (base pairs) are indicated to the left of the panel. B. Graphical representation of the bbb26–27 region on cp26 (B31-A) and the cloned pieces of DNA used for the allelic exchange constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ). The 1168 bp region of cp26 between nucleotides 21923 and 23091 was replaced with the 1146 bp flaB p – aadA resistance cassette to create XL-BBB26–27Δ for disruption of both bbb26 and bbb27 . The 742 bp region of cp26 between nucleotides 21923 and 22579 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB26Δ for disruption of bbb26 . The 438 bp region of cp26 between nucleotides 22653 and 23091 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB27Δ for disruption of bbb27 . Locations of the primers used for analysis in (A) are indicated and the sequences are listed in Table S1 .

    Journal: Molecular Microbiology

    Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi

    doi: 10.1111/j.1365-2958.2007.05969.x

    Figure Lengend Snippet: A. PCR analysis of genomic DNA from B. burgdorferi clones transformed with gene inactivation constructs targeting genes in the bbb26–27 region. Template DNAs from transformants are identified below the lanes and PCR amplification targets above the lanes. Template DNA from B31-A illustrates the PCR products from the wild-type alleles of bbb26–27 , bbb26 and bbb27 (lanes 2–5, 15 and 19), whereas template DNAs from the gene inactivation constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ) depict the PCR profiles of the mutated alleles (lanes 6–9, 16 and 20). The PCR products resulting from the clones transformed with the allelic exchange inactivation plasmids are illustrated in lanes 10–13, 17 and 21. The 1 kbp-plus size standards (Invitrogen) were run in lanes 1, 14 and 18 and sizes (base pairs) are indicated to the left of the panel. B. Graphical representation of the bbb26–27 region on cp26 (B31-A) and the cloned pieces of DNA used for the allelic exchange constructs (XL-BBB26–27Δ, XL-BBB26Δ and XL-BBB27Δ). The 1168 bp region of cp26 between nucleotides 21923 and 23091 was replaced with the 1146 bp flaB p – aadA resistance cassette to create XL-BBB26–27Δ for disruption of both bbb26 and bbb27 . The 742 bp region of cp26 between nucleotides 21923 and 22579 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB26Δ for disruption of bbb26 . The 438 bp region of cp26 between nucleotides 22653 and 23091 was replaced with the 1100 bp flgB p – aacC1 resistance cassette to create XL-BBB27Δ for disruption of bbb27 . Locations of the primers used for analysis in (A) are indicated and the sequences are listed in Table S1 .

    Article Snippet: B31 clone A (B31-A) ( ) is a non-infectious derivative of type strain B31 (ATCC 35210), which was isolated from a tick collected on Shelter Island in New York ( ).

    Techniques: Polymerase Chain Reaction, Clone Assay, Transformation Assay, Construct, Amplification

    Evaluation of untreated (control) and treated with Baicalein (500 μ g ml −1 ) or Monolaurin (500 μ g ml −1 ) biofilm of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h as a qualitative measurement using LIVE/DEAD BacLight staining; green – live biofilm, red – dead biofilm, images taken at 4 × magnification. Immunofluorescence staining with primary antibodies against untreated B. burgdorferi B31 strain (c) and B. garinii CIP103362 strain (d) after 72 h using AlexaFluor‐488 as a secondary antibody respectively; images taken at 10× (right) and 63× (left) magnification.

    Journal: Journal of Applied Microbiology

    Article Title: In vitro evaluation of antibacterial activity of phytochemicals and micronutrients against Borrelia burgdorferi and Borrelia garinii

    doi: 10.1111/jam.12970

    Figure Lengend Snippet: Evaluation of untreated (control) and treated with Baicalein (500 μ g ml −1 ) or Monolaurin (500 μ g ml −1 ) biofilm of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h as a qualitative measurement using LIVE/DEAD BacLight staining; green – live biofilm, red – dead biofilm, images taken at 4 × magnification. Immunofluorescence staining with primary antibodies against untreated B. burgdorferi B31 strain (c) and B. garinii CIP103362 strain (d) after 72 h using AlexaFluor‐488 as a secondary antibody respectively; images taken at 10× (right) and 63× (left) magnification.

    Article Snippet: Low passage isolates of the B31 strain of B. burgdorferi and CIP103362 strain of B. garinii were obtained from the American Type Culture Collection (Manassas, VA).

    Techniques: Staining, Immunofluorescence

    Kinetic evaluation of growth of live/dead untreated spirochetes of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) monitored up to 72 h. Live spirochetes (■), dead spirochetes ( ), * !blank; P ≤ 0·001.

    Journal: Journal of Applied Microbiology

    Article Title: In vitro evaluation of antibacterial activity of phytochemicals and micronutrients against Borrelia burgdorferi and Borrelia garinii

    doi: 10.1111/jam.12970

    Figure Lengend Snippet: Kinetic evaluation of growth of live/dead untreated spirochetes of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) monitored up to 72 h. Live spirochetes (■), dead spirochetes ( ), * !blank; P ≤ 0·001.

    Article Snippet: Low passage isolates of the B31 strain of B. burgdorferi and CIP103362 strain of B. garinii were obtained from the American Type Culture Collection (Manassas, VA).

    Techniques:

    Evaluation of untreated (control) and treated with Baicalein (350 μ g ml −1 ) or Monolaurin (300 μ g ml −1 ) Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h as a qualitative measurement by fluorescence microscope using LIVE/DEAD BacLight staining; green fluorescence – live organisms (spirochetes and rounded forms), red fluorescence – dead organisms (spirochetes and rounded forms). Merged images taken at 20× magnification.

    Journal: Journal of Applied Microbiology

    Article Title: In vitro evaluation of antibacterial activity of phytochemicals and micronutrients against Borrelia burgdorferi and Borrelia garinii

    doi: 10.1111/jam.12970

    Figure Lengend Snippet: Evaluation of untreated (control) and treated with Baicalein (350 μ g ml −1 ) or Monolaurin (300 μ g ml −1 ) Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h as a qualitative measurement by fluorescence microscope using LIVE/DEAD BacLight staining; green fluorescence – live organisms (spirochetes and rounded forms), red fluorescence – dead organisms (spirochetes and rounded forms). Merged images taken at 20× magnification.

    Article Snippet: Low passage isolates of the B31 strain of B. burgdorferi and CIP103362 strain of B. garinii were obtained from the American Type Culture Collection (Manassas, VA).

    Techniques: Fluorescence, Microscopy, Staining

    Kinetic evaluation of bactericidal effect of phytochemicals against the spirochetes (a and b) and rounded forms (c and d) of Borrelia burgdorferi B31 strain (a and c) and Borrelia garinii CIP103362 strain (b and d) monitored up to 72 h. (a and b) All data are statistically significant; tested compounds: 250 μ g ml −1 Baicalein (▼), 250 μ g ml −1 Monolaurin (∆), 250 μ g ml −1 Cis‐2‐decenoic acid (●), 20 μ g ml −1 Kelp (Iodine) (□). (c and d) LD50 marked with the solid line; # P ≤ 0·05, ∆ P ≤ 0·01, * P ≤ 0·001; tested compounds: 350 μ g ml −1 Baicalein, 300 μ g ml −1 Monolaurin, 500 μ g ml −1 Cis‐2‐decenoic acid, 20 μ g ml −1 Kelp (Iodine); 24 h (■), 48 h ( ), 72 h ( ).

    Journal: Journal of Applied Microbiology

    Article Title: In vitro evaluation of antibacterial activity of phytochemicals and micronutrients against Borrelia burgdorferi and Borrelia garinii

    doi: 10.1111/jam.12970

    Figure Lengend Snippet: Kinetic evaluation of bactericidal effect of phytochemicals against the spirochetes (a and b) and rounded forms (c and d) of Borrelia burgdorferi B31 strain (a and c) and Borrelia garinii CIP103362 strain (b and d) monitored up to 72 h. (a and b) All data are statistically significant; tested compounds: 250 μ g ml −1 Baicalein (▼), 250 μ g ml −1 Monolaurin (∆), 250 μ g ml −1 Cis‐2‐decenoic acid (●), 20 μ g ml −1 Kelp (Iodine) (□). (c and d) LD50 marked with the solid line; # P ≤ 0·05, ∆ P ≤ 0·01, * P ≤ 0·001; tested compounds: 350 μ g ml −1 Baicalein, 300 μ g ml −1 Monolaurin, 500 μ g ml −1 Cis‐2‐decenoic acid, 20 μ g ml −1 Kelp (Iodine); 24 h (■), 48 h ( ), 72 h ( ).

    Article Snippet: Low passage isolates of the B31 strain of B. burgdorferi and CIP103362 strain of B. garinii were obtained from the American Type Culture Collection (Manassas, VA).

    Techniques:

    Dose‐dependent estimation of remaining biofilm of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h of post‐treatment with phytochemicals and antibiotic doxycycline presented as a quantitative examination; # P ≤ 0·05, ∆ P ≤ 0·01.

    Journal: Journal of Applied Microbiology

    Article Title: In vitro evaluation of antibacterial activity of phytochemicals and micronutrients against Borrelia burgdorferi and Borrelia garinii

    doi: 10.1111/jam.12970

    Figure Lengend Snippet: Dose‐dependent estimation of remaining biofilm of Borrelia burgdorferi B31 strain (a) and Borrelia garinii CIP103362 strain (b) after 72 h of post‐treatment with phytochemicals and antibiotic doxycycline presented as a quantitative examination; # P ≤ 0·05, ∆ P ≤ 0·01.

    Article Snippet: Low passage isolates of the B31 strain of B. burgdorferi and CIP103362 strain of B. garinii were obtained from the American Type Culture Collection (Manassas, VA).

    Techniques:

    OspC production by WT and A3/ flaB p :: ospC spirochetes in unfed nymphs. Nymphs were artificially infected as larvae with wild-type B. burgdorferi strain B31-A3 (WT) or an isogenic derivative engineered to constitutively express ospC ( A3/ flaB p :: ospC ). Spirochetes in dissected tick midguts were detected by IFA with a polyclonal anti- B. burgdorferi primary antiserum [ 41 ] and rhodamine-labeled secondary antibody, while synthesis of OspC was examined using a monoclonal anti-OspC primary antibody [ 42 ] and FITC-labeled secondary antibody

    Journal: Parasites & Vectors

    Article Title: Virulence of the Lyme disease spirochete before and after the tick bloodmeal: a quantitative assessment

    doi: 10.1186/s13071-016-1380-1

    Figure Lengend Snippet: OspC production by WT and A3/ flaB p :: ospC spirochetes in unfed nymphs. Nymphs were artificially infected as larvae with wild-type B. burgdorferi strain B31-A3 (WT) or an isogenic derivative engineered to constitutively express ospC ( A3/ flaB p :: ospC ). Spirochetes in dissected tick midguts were detected by IFA with a polyclonal anti- B. burgdorferi primary antiserum [ 41 ] and rhodamine-labeled secondary antibody, while synthesis of OspC was examined using a monoclonal anti-OspC primary antibody [ 42 ] and FITC-labeled secondary antibody

    Article Snippet: Borrelia burgdorferi strains and culture conditions Infectious clone B31-A3, derived from the B31 type strain of B. burgdorferi (ATCC 35210), was used as the wild type (WT) strain, and an isogenic derivative carrying the ospC gene driven by the constitutive flaB promoter on the shuttle vector pBSV2G, termed A3/flaB p ::ospC , was used for the constitutive ospC expression experiments [ , ].

    Techniques: Infection, Immunofluorescence, Labeling

    Cross-reacting Bdr paralog and ortholog proteins in other Borrelia isolates. (A) Coomassie blue-stained SDS–12% polyacrylamide gel. Borrelia isolates are indicated above the lanes: m, molecular weight protein marker (Gibco high molecular weight); B. burgdorferi sensu stricto isolates B31, B313, Sh-2-82, N40, and HB19; B. garinii Ip90; B. afzelii ACA1; tick-borne relapsing fever isolates B. turicatae Oz1 (Bt) and B. hermsii HS1 (Bh). (B) Western immunoblot of an identical gel with anti-BdrA antiserum (1:500). Lanes are as in panel A.

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: Cross-reacting Bdr paralog and ortholog proteins in other Borrelia isolates. (A) Coomassie blue-stained SDS–12% polyacrylamide gel. Borrelia isolates are indicated above the lanes: m, molecular weight protein marker (Gibco high molecular weight); B. burgdorferi sensu stricto isolates B31, B313, Sh-2-82, N40, and HB19; B. garinii Ip90; B. afzelii ACA1; tick-borne relapsing fever isolates B. turicatae Oz1 (Bt) and B. hermsii HS1 (Bh). (B) Western immunoblot of an identical gel with anti-BdrA antiserum (1:500). Lanes are as in panel A.

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Staining, Molecular Weight, Marker, Western Blot

    Heterologous expression and cross-reactivity of B31 Bdr proteins. (A) Coomassie blue-stained SDS–12% polyacrylamide gel. Lanes: m, molecular weight marker (Gibco high molecular weight); 1, B. burgdorferi B31 whole-cell lysate; 2, E. coli BL21(DE3)(pLysS)(pET29b) whole-cell lysate; 3, E. coli BL21(DE3)(pLysS)(pWRZ102) whole-cell lysate (expressing rBdrV); 4, E. coli BL21(DE3)(pLysS)(pWRZ101) whole-cell lysate (expressing rBdrA); 5, purified rBdrA. Asterisks indicate rBdrV and rBdrA. (B) Western immunoblot with anti-BdrA, anti-BdrV, and control rabbit sera (1:500). Lanes are as in panel A.

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: Heterologous expression and cross-reactivity of B31 Bdr proteins. (A) Coomassie blue-stained SDS–12% polyacrylamide gel. Lanes: m, molecular weight marker (Gibco high molecular weight); 1, B. burgdorferi B31 whole-cell lysate; 2, E. coli BL21(DE3)(pLysS)(pET29b) whole-cell lysate; 3, E. coli BL21(DE3)(pLysS)(pWRZ102) whole-cell lysate (expressing rBdrV); 4, E. coli BL21(DE3)(pLysS)(pWRZ101) whole-cell lysate (expressing rBdrA); 5, purified rBdrA. Asterisks indicate rBdrV and rBdrA. (B) Western immunoblot with anti-BdrA, anti-BdrV, and control rabbit sera (1:500). Lanes are as in panel A.

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Expressing, Staining, Molecular Weight, Marker, Purification, Western Blot

    Lack of temperature-dependent expression and proteinase K susceptibility of Bdr proteins. (A) Western immunoblot of an SDS–12% polyacrylamide gel with whole-cell lysates of B. burgdorferi B31 grown at 24 or 37°C probed with anti-BdrA antiserum (1:500). Protein sizes indicated to the left are derived from a molecular weight marker (Gibco high molecular weight). (B) Proteinase K susceptibility of Bdr proteins. Coom. Blue, Coomassie blue-stained SDS–12% polyacrylamide gel with whole-cell lysates of B. burgdorferi B31 either untreated (−) or previously treated (+) with proteinase K (200 μg/ml of cell suspension); anti-BrdA and anti-OspA, Western immunoblots of gels with anti-BdrA (1:500) and anti-OspA (1:20,000) antisera. An asterisk indicates OspA.

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: Lack of temperature-dependent expression and proteinase K susceptibility of Bdr proteins. (A) Western immunoblot of an SDS–12% polyacrylamide gel with whole-cell lysates of B. burgdorferi B31 grown at 24 or 37°C probed with anti-BdrA antiserum (1:500). Protein sizes indicated to the left are derived from a molecular weight marker (Gibco high molecular weight). (B) Proteinase K susceptibility of Bdr proteins. Coom. Blue, Coomassie blue-stained SDS–12% polyacrylamide gel with whole-cell lysates of B. burgdorferi B31 either untreated (−) or previously treated (+) with proteinase K (200 μg/ml of cell suspension); anti-BrdA and anti-OspA, Western immunoblots of gels with anti-BdrA (1:500) and anti-OspA (1:20,000) antisera. An asterisk indicates OspA.

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Expressing, Western Blot, Derivative Assay, Molecular Weight, Marker, Staining

    Immunofluorescence microscopy of fixed and unfixed B. burgdorferi cells. (A) Methanol-fixed, permeabilized B. burgdorferi B31 cells reacted with anti-OspA (1:1,000), anti-BdrA (1:200), and control (1:200) polyclonal rabbit sera. Secondary antibody was a fluorescein isothiocyanate-labeled goat anti-rabbit antibody. (B) Unfixed, intact B. burgdorferi B31 cells reacted with anti-OspA (1:1,000), anti-BdrA (1:100), and control (1:100) polyclonal rabbit sera.

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: Immunofluorescence microscopy of fixed and unfixed B. burgdorferi cells. (A) Methanol-fixed, permeabilized B. burgdorferi B31 cells reacted with anti-OspA (1:1,000), anti-BdrA (1:200), and control (1:200) polyclonal rabbit sera. Secondary antibody was a fluorescein isothiocyanate-labeled goat anti-rabbit antibody. (B) Unfixed, intact B. burgdorferi B31 cells reacted with anti-OspA (1:1,000), anti-BdrA (1:100), and control (1:100) polyclonal rabbit sera.

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Immunofluorescence, Microscopy, Labeling

    Western immunoblots and ELISA with Lyme disease patient sera. (A) Left, Western immunoblot with a representative positive Lyme disease (LD) patient serum (1:100). Lane 1, B. burgdorferi B31 whole-cell lysate; lane 2, E. coli BL21(DE3)(pLysS)(pET29b) whole-cell lysate; lane 3, E. coli BL21(DE3)(pLysS)(pWRZ102) whole-cell lysate (expressing rBdrV); lane 4, E. coli BL21(DE3)(pLysS)(pWRZ101) whole-cell lysate (expressing rBdrA); lane 5, purified rBdrA. Right, Western immunoblot of an identical blot with a representative human control serum. (B) Distribution of rounded ELISA OD values obtained with LD patient and control sera. A triangle on the x axis indicates the cutoff OD (mean of control sera values plus 3 SD = 0.20) for positive sera.

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: Western immunoblots and ELISA with Lyme disease patient sera. (A) Left, Western immunoblot with a representative positive Lyme disease (LD) patient serum (1:100). Lane 1, B. burgdorferi B31 whole-cell lysate; lane 2, E. coli BL21(DE3)(pLysS)(pET29b) whole-cell lysate; lane 3, E. coli BL21(DE3)(pLysS)(pWRZ102) whole-cell lysate (expressing rBdrV); lane 4, E. coli BL21(DE3)(pLysS)(pWRZ101) whole-cell lysate (expressing rBdrA); lane 5, purified rBdrA. Right, Western immunoblot of an identical blot with a representative human control serum. (B) Distribution of rounded ELISA OD values obtained with LD patient and control sera. A triangle on the x axis indicates the cutoff OD (mean of control sera values plus 3 SD = 0.20) for positive sera.

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Purification

    cp32 plasmid profile of B. burgdorferi B31 derivative B313. Plasmid-enriched DNA of B. burgdorferi B31 and its derivative B313 was dot blotted with nucleotide probes (1 through 9) specific for the multiple homologous 32-kb-long circular plasmids (cp32-1 through cp32-9). Probes were also blotted as a control for probe specificity. Note that probe 2/7 does not discriminate between cp32-2 and cp32-7. PCR analysis with oligonucleotide primers specific for cp32-2- or cp32-7 revealed that both B31 and B313 carried cp32-2 but not cp32-7 (see text).

    Journal: Infection and Immunity

    Article Title: Comparative Analysis and Immunological Characterization of the Borrelia Bdr Protein Family

    doi:

    Figure Lengend Snippet: cp32 plasmid profile of B. burgdorferi B31 derivative B313. Plasmid-enriched DNA of B. burgdorferi B31 and its derivative B313 was dot blotted with nucleotide probes (1 through 9) specific for the multiple homologous 32-kb-long circular plasmids (cp32-1 through cp32-9). Probes were also blotted as a control for probe specificity. Note that probe 2/7 does not discriminate between cp32-2 and cp32-7. PCR analysis with oligonucleotide primers specific for cp32-2- or cp32-7 revealed that both B31 and B313 carried cp32-2 but not cp32-7 (see text).

    Article Snippet: Including the two initially identified copies on one of the multiple homologous cp32s and the related lp56 of B. burgdorferi B31 (ORF-E [ ]), a total of 29 related sequences have been determined, mostly as part of the B. burgdorferi B31 genome sequencing project but also in other B. burgdorferi sensu lato strains and the relapsing fever spirochetes B. hermsii and B. turicatae (Table ).

    Techniques: Plasmid Preparation, Polymerase Chain Reaction