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John F. Bohnsack - One of the best experts on this subject based on the ideXlab platform.
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Long-range mapping of the Streptococcus agalactiae phylogenetic lineage Restriction Digest pattern type III-3 reveals clustering of virulence genes.
Infection and immunity, 2002Co-Authors: John F. Bohnsack, April Whiting, Russell D. Bradford, Brenna K. Van Frank, Shinji Takahashi, Elisabeth E. AddersonAbstract:Human isolates of serotype III Streptococcus agalactiae (group B streptococcus [GBS]) can be divided into three separate phylogenetic lineages based on analysis of the Restriction Digest patterns (RDPs) of chromosomal DNA. Nine DNA sequences that are present in all isolates of the RDP III-3 phylogenetic lineage, but not in the other lineages, were identified by genomic subtractive hybridization. A complete physical map of a III-3 chromosome was constructed. Six of the nine III-3-specific sequences mapped to a 340-kb Sse8387I fragment which contains or is located close to known GBS virulence genes. One of the III-3-specific probes, AW-10, encodes part of GBSi1, a group II intron that is inserted at two sites within the GBS genome. The second chromosomal site for GBSi1 was isolated, sequenced, and mapped to a location near the locus responsible for hemolysin production. These findings suggest that the genetic variation that distinguishes the RDP type III-3 strains from other serotype III strains occurs largely within localized areas of the genome containing known or putative virulence genes.
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Phylogenetic Classification of Serotype III Group B Streptococci on the Basis of hylB Gene Analysis and DNA Sequences Specific to Restriction Digest Pattern Type III-3
The Journal of infectious diseases, 2001Co-Authors: John F. Bohnsack, April Whiting, Shinji Takahashi, Shauna Detrick, Leslie R. Pelinka, Laura L. Hammitt, Adrienne A. Aly, Elisabeth E. AddersonAbstract:Previous work divided serotype III group B streptococci (GBS) into 3 major phylogenetic lineages (III-1, III-2, and III-3) on the basis of bacterial DNA Restriction Digest patterns (RDPs). Most neonatal invasive disease was caused by III-3 strains, which implies that III-3 strains are more virulent than III-2 or III-1 strains. In the current studies, all RDP III-3 and III-1 strains expressed hyaluronate lysase activity; however, all III-2 strains lack hyaluronate lysase activity, because the gene that encodes hyaluronate lysase, hylB, is inactivated by IS1548. Subtractive hybridization was used to identify 9 short DNA sequences that are present in all the III-3 strains but not in any of the III-2 or III-1 strains. With 1 exception, these III-3-specific sequences were not detected in nonserotype III GBS. These data further validate the RDP-based subclassification of GBS and suggest that lineage-specific genes will be identified, which account for the differences in virulence among the lineages.
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correlation of hind iii Restriction Digest patterns of chromosomal dna from type iii group b streptococci with invasive disease 1101
Pediatric Research, 1996Co-Authors: Shinji Takahashi, Mark R. Briesacher, Judy A. Daly, Karen C. Carroll, Harry R. Hill, John F. BohnsackAbstract:Aside from the polysaccharide capsule, little is known about bacterial factors that contribute to the virulence of group B streptococci (GBS). In an effort to identify bacteria more likely to cause invasive disease, we determined GBS C5a-ase activity and HindIII-derived Restriction Digest patterns (RDPs) of chromosomal DNA from 41 invasive type III GBS strains isolated from sterile body sites (blood, CSF, or joint) and 29 colonizing strains isolated from vaginal secretions. The similarity between RDPs was expressed as a Pearson product moment coefficient that compares densitometric analyses of RDPs between strains (J. Med. Microbiol. 35: 297, 1991). Bacteria were grouped into one of four RDP types: III-1 (4.3%), III-2(25.7%), III-3 (58.6%) and III-4 (11.4%). Bacterial RDPs within each type were similar at a coefficient value ≥0.93. There was no obvious correlation between expression of C5a-ase and invasiveness in any of the RDP types. All type III-1 and III-2 strains tested expressed C5a-ase, while all of the type III-4 strains did not express C5a-ase activity. 66% of the III-3 strains expressed C5a-ase, but C5a-ase negative and positive strains were equally likely to be invasive isolates. Type III-3 strains that were very similar to each other by RDP typing (coefficient ≥0.99) were concordant for the presence or absence of C5a-ase, supporting the assertion that the RDP typing identifies closely related strains. Invasive isolates were significantly more likely to be type III-3 (78%) than were colonizing isolates (33%) (p<.001, Chi-squared). These data suggest that RDP type III-3 strains are more likely to cause invasive disease in human infants than other RDP types. Further characterization of these strains may help identify type III GBS virulence factors.
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CORRELATION OF HIND III Restriction Digest PATTERNS OF CHROMOSOMAL DNA FROM TYPE III GROUP B STREPTOCOCCI WITH INVASIVE DISEASE. • 1101
Pediatric Research, 1996Co-Authors: Shinji Takahashi, Mark R. Briesacher, Judy A. Daly, Karen C. Carroll, Harry R. Hill, John F. BohnsackAbstract:Aside from the polysaccharide capsule, little is known about bacterial factors that contribute to the virulence of group B streptococci (GBS). In an effort to identify bacteria more likely to cause invasive disease, we determined GBS C5a-ase activity and HindIII-derived Restriction Digest patterns (RDPs) of chromosomal DNA from 41 invasive type III GBS strains isolated from sterile body sites (blood, CSF, or joint) and 29 colonizing strains isolated from vaginal secretions. The similarity between RDPs was expressed as a Pearson product moment coefficient that compares densitometric analyses of RDPs between strains (J. Med. Microbiol. 35: 297, 1991). Bacteria were grouped into one of four RDP types: III-1 (4.3%), III-2(25.7%), III-3 (58.6%) and III-4 (11.4%). Bacterial RDPs within each type were similar at a coefficient value ≥0.93. There was no obvious correlation between expression of C5a-ase and invasiveness in any of the RDP types. All type III-1 and III-2 strains tested expressed C5a-ase, while all of the type III-4 strains did not express C5a-ase activity. 66% of the III-3 strains expressed C5a-ase, but C5a-ase negative and positive strains were equally likely to be invasive isolates. Type III-3 strains that were very similar to each other by RDP typing (coefficient ≥0.99) were concordant for the presence or absence of C5a-ase, supporting the assertion that the RDP typing identifies closely related strains. Invasive isolates were significantly more likely to be type III-3 (78%) than were colonizing isolates (33%) (p
Shinji Takahashi - One of the best experts on this subject based on the ideXlab platform.
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Long-range mapping of the Streptococcus agalactiae phylogenetic lineage Restriction Digest pattern type III-3 reveals clustering of virulence genes.
Infection and immunity, 2002Co-Authors: John F. Bohnsack, April Whiting, Russell D. Bradford, Brenna K. Van Frank, Shinji Takahashi, Elisabeth E. AddersonAbstract:Human isolates of serotype III Streptococcus agalactiae (group B streptococcus [GBS]) can be divided into three separate phylogenetic lineages based on analysis of the Restriction Digest patterns (RDPs) of chromosomal DNA. Nine DNA sequences that are present in all isolates of the RDP III-3 phylogenetic lineage, but not in the other lineages, were identified by genomic subtractive hybridization. A complete physical map of a III-3 chromosome was constructed. Six of the nine III-3-specific sequences mapped to a 340-kb Sse8387I fragment which contains or is located close to known GBS virulence genes. One of the III-3-specific probes, AW-10, encodes part of GBSi1, a group II intron that is inserted at two sites within the GBS genome. The second chromosomal site for GBSi1 was isolated, sequenced, and mapped to a location near the locus responsible for hemolysin production. These findings suggest that the genetic variation that distinguishes the RDP type III-3 strains from other serotype III strains occurs largely within localized areas of the genome containing known or putative virulence genes.
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Phylogenetic Classification of Serotype III Group B Streptococci on the Basis of hylB Gene Analysis and DNA Sequences Specific to Restriction Digest Pattern Type III-3
The Journal of infectious diseases, 2001Co-Authors: John F. Bohnsack, April Whiting, Shinji Takahashi, Shauna Detrick, Leslie R. Pelinka, Laura L. Hammitt, Adrienne A. Aly, Elisabeth E. AddersonAbstract:Previous work divided serotype III group B streptococci (GBS) into 3 major phylogenetic lineages (III-1, III-2, and III-3) on the basis of bacterial DNA Restriction Digest patterns (RDPs). Most neonatal invasive disease was caused by III-3 strains, which implies that III-3 strains are more virulent than III-2 or III-1 strains. In the current studies, all RDP III-3 and III-1 strains expressed hyaluronate lysase activity; however, all III-2 strains lack hyaluronate lysase activity, because the gene that encodes hyaluronate lysase, hylB, is inactivated by IS1548. Subtractive hybridization was used to identify 9 short DNA sequences that are present in all the III-3 strains but not in any of the III-2 or III-1 strains. With 1 exception, these III-3-specific sequences were not detected in nonserotype III GBS. These data further validate the RDP-based subclassification of GBS and suggest that lineage-specific genes will be identified, which account for the differences in virulence among the lineages.
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correlation of hind iii Restriction Digest patterns of chromosomal dna from type iii group b streptococci with invasive disease 1101
Pediatric Research, 1996Co-Authors: Shinji Takahashi, Mark R. Briesacher, Judy A. Daly, Karen C. Carroll, Harry R. Hill, John F. BohnsackAbstract:Aside from the polysaccharide capsule, little is known about bacterial factors that contribute to the virulence of group B streptococci (GBS). In an effort to identify bacteria more likely to cause invasive disease, we determined GBS C5a-ase activity and HindIII-derived Restriction Digest patterns (RDPs) of chromosomal DNA from 41 invasive type III GBS strains isolated from sterile body sites (blood, CSF, or joint) and 29 colonizing strains isolated from vaginal secretions. The similarity between RDPs was expressed as a Pearson product moment coefficient that compares densitometric analyses of RDPs between strains (J. Med. Microbiol. 35: 297, 1991). Bacteria were grouped into one of four RDP types: III-1 (4.3%), III-2(25.7%), III-3 (58.6%) and III-4 (11.4%). Bacterial RDPs within each type were similar at a coefficient value ≥0.93. There was no obvious correlation between expression of C5a-ase and invasiveness in any of the RDP types. All type III-1 and III-2 strains tested expressed C5a-ase, while all of the type III-4 strains did not express C5a-ase activity. 66% of the III-3 strains expressed C5a-ase, but C5a-ase negative and positive strains were equally likely to be invasive isolates. Type III-3 strains that were very similar to each other by RDP typing (coefficient ≥0.99) were concordant for the presence or absence of C5a-ase, supporting the assertion that the RDP typing identifies closely related strains. Invasive isolates were significantly more likely to be type III-3 (78%) than were colonizing isolates (33%) (p<.001, Chi-squared). These data suggest that RDP type III-3 strains are more likely to cause invasive disease in human infants than other RDP types. Further characterization of these strains may help identify type III GBS virulence factors.
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CORRELATION OF HIND III Restriction Digest PATTERNS OF CHROMOSOMAL DNA FROM TYPE III GROUP B STREPTOCOCCI WITH INVASIVE DISEASE. • 1101
Pediatric Research, 1996Co-Authors: Shinji Takahashi, Mark R. Briesacher, Judy A. Daly, Karen C. Carroll, Harry R. Hill, John F. BohnsackAbstract:Aside from the polysaccharide capsule, little is known about bacterial factors that contribute to the virulence of group B streptococci (GBS). In an effort to identify bacteria more likely to cause invasive disease, we determined GBS C5a-ase activity and HindIII-derived Restriction Digest patterns (RDPs) of chromosomal DNA from 41 invasive type III GBS strains isolated from sterile body sites (blood, CSF, or joint) and 29 colonizing strains isolated from vaginal secretions. The similarity between RDPs was expressed as a Pearson product moment coefficient that compares densitometric analyses of RDPs between strains (J. Med. Microbiol. 35: 297, 1991). Bacteria were grouped into one of four RDP types: III-1 (4.3%), III-2(25.7%), III-3 (58.6%) and III-4 (11.4%). Bacterial RDPs within each type were similar at a coefficient value ≥0.93. There was no obvious correlation between expression of C5a-ase and invasiveness in any of the RDP types. All type III-1 and III-2 strains tested expressed C5a-ase, while all of the type III-4 strains did not express C5a-ase activity. 66% of the III-3 strains expressed C5a-ase, but C5a-ase negative and positive strains were equally likely to be invasive isolates. Type III-3 strains that were very similar to each other by RDP typing (coefficient ≥0.99) were concordant for the presence or absence of C5a-ase, supporting the assertion that the RDP typing identifies closely related strains. Invasive isolates were significantly more likely to be type III-3 (78%) than were colonizing isolates (33%) (p
Elisabeth E. Adderson - One of the best experts on this subject based on the ideXlab platform.
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Long-range mapping of the Streptococcus agalactiae phylogenetic lineage Restriction Digest pattern type III-3 reveals clustering of virulence genes.
Infection and immunity, 2002Co-Authors: John F. Bohnsack, April Whiting, Russell D. Bradford, Brenna K. Van Frank, Shinji Takahashi, Elisabeth E. AddersonAbstract:Human isolates of serotype III Streptococcus agalactiae (group B streptococcus [GBS]) can be divided into three separate phylogenetic lineages based on analysis of the Restriction Digest patterns (RDPs) of chromosomal DNA. Nine DNA sequences that are present in all isolates of the RDP III-3 phylogenetic lineage, but not in the other lineages, were identified by genomic subtractive hybridization. A complete physical map of a III-3 chromosome was constructed. Six of the nine III-3-specific sequences mapped to a 340-kb Sse8387I fragment which contains or is located close to known GBS virulence genes. One of the III-3-specific probes, AW-10, encodes part of GBSi1, a group II intron that is inserted at two sites within the GBS genome. The second chromosomal site for GBSi1 was isolated, sequenced, and mapped to a location near the locus responsible for hemolysin production. These findings suggest that the genetic variation that distinguishes the RDP type III-3 strains from other serotype III strains occurs largely within localized areas of the genome containing known or putative virulence genes.
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Phylogenetic Classification of Serotype III Group B Streptococci on the Basis of hylB Gene Analysis and DNA Sequences Specific to Restriction Digest Pattern Type III-3
The Journal of infectious diseases, 2001Co-Authors: John F. Bohnsack, April Whiting, Shinji Takahashi, Shauna Detrick, Leslie R. Pelinka, Laura L. Hammitt, Adrienne A. Aly, Elisabeth E. AddersonAbstract:Previous work divided serotype III group B streptococci (GBS) into 3 major phylogenetic lineages (III-1, III-2, and III-3) on the basis of bacterial DNA Restriction Digest patterns (RDPs). Most neonatal invasive disease was caused by III-3 strains, which implies that III-3 strains are more virulent than III-2 or III-1 strains. In the current studies, all RDP III-3 and III-1 strains expressed hyaluronate lysase activity; however, all III-2 strains lack hyaluronate lysase activity, because the gene that encodes hyaluronate lysase, hylB, is inactivated by IS1548. Subtractive hybridization was used to identify 9 short DNA sequences that are present in all the III-3 strains but not in any of the III-2 or III-1 strains. With 1 exception, these III-3-specific sequences were not detected in nonserotype III GBS. These data further validate the RDP-based subclassification of GBS and suggest that lineage-specific genes will be identified, which account for the differences in virulence among the lineages.
April Whiting - One of the best experts on this subject based on the ideXlab platform.
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Long-range mapping of the Streptococcus agalactiae phylogenetic lineage Restriction Digest pattern type III-3 reveals clustering of virulence genes.
Infection and immunity, 2002Co-Authors: John F. Bohnsack, April Whiting, Russell D. Bradford, Brenna K. Van Frank, Shinji Takahashi, Elisabeth E. AddersonAbstract:Human isolates of serotype III Streptococcus agalactiae (group B streptococcus [GBS]) can be divided into three separate phylogenetic lineages based on analysis of the Restriction Digest patterns (RDPs) of chromosomal DNA. Nine DNA sequences that are present in all isolates of the RDP III-3 phylogenetic lineage, but not in the other lineages, were identified by genomic subtractive hybridization. A complete physical map of a III-3 chromosome was constructed. Six of the nine III-3-specific sequences mapped to a 340-kb Sse8387I fragment which contains or is located close to known GBS virulence genes. One of the III-3-specific probes, AW-10, encodes part of GBSi1, a group II intron that is inserted at two sites within the GBS genome. The second chromosomal site for GBSi1 was isolated, sequenced, and mapped to a location near the locus responsible for hemolysin production. These findings suggest that the genetic variation that distinguishes the RDP type III-3 strains from other serotype III strains occurs largely within localized areas of the genome containing known or putative virulence genes.
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Phylogenetic Classification of Serotype III Group B Streptococci on the Basis of hylB Gene Analysis and DNA Sequences Specific to Restriction Digest Pattern Type III-3
The Journal of infectious diseases, 2001Co-Authors: John F. Bohnsack, April Whiting, Shinji Takahashi, Shauna Detrick, Leslie R. Pelinka, Laura L. Hammitt, Adrienne A. Aly, Elisabeth E. AddersonAbstract:Previous work divided serotype III group B streptococci (GBS) into 3 major phylogenetic lineages (III-1, III-2, and III-3) on the basis of bacterial DNA Restriction Digest patterns (RDPs). Most neonatal invasive disease was caused by III-3 strains, which implies that III-3 strains are more virulent than III-2 or III-1 strains. In the current studies, all RDP III-3 and III-1 strains expressed hyaluronate lysase activity; however, all III-2 strains lack hyaluronate lysase activity, because the gene that encodes hyaluronate lysase, hylB, is inactivated by IS1548. Subtractive hybridization was used to identify 9 short DNA sequences that are present in all the III-3 strains but not in any of the III-2 or III-1 strains. With 1 exception, these III-3-specific sequences were not detected in nonserotype III GBS. These data further validate the RDP-based subclassification of GBS and suggest that lineage-specific genes will be identified, which account for the differences in virulence among the lineages.
Marco A Marra - One of the best experts on this subject based on the ideXlab platform.
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large scale bac clone Restriction Digest fingerprinting
Current protocols in human genetics, 2007Co-Authors: Carrie Mathewson, Jacqueline E Schein, Marco A MarraAbstract:Restriction Digest fingerprinting is a common method for characterizing large insert genomic clones, e.g., bacterial artificial chromosome (BAC), P1 artificial chromosome (PAC) and Fosmid clones. This clone fingerprinting method has been widely applied in the construction of clone-based physical maps, which have been used as positional cloning resources as well as to support directed and genome-wide sequencing efforts. This unit describes a robust, large-scale procedure for generation of agarose gel–based clone fingerprints from BAC clones. Keywords: Restriction Digest fingerprint; bacterial artificial chromosome (BAC); BAC DNA isolation; agarose gel electrophoresis; BAC fingerprint map
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Current Protocols in Human Genetics - Large‐Scale BAC Clone Restriction Digest Fingerprinting
Current Protocols in Human Genetics, 2007Co-Authors: Carrie Mathewson, Jacqueline E Schein, Marco A MarraAbstract:Restriction Digest fingerprinting is a common method for characterizing large insert genomic clones, e.g., bacterial artificial chromosome (BAC), P1 artificial chromosome (PAC) and Fosmid clones. This clone fingerprinting method has been widely applied in the construction of clone-based physical maps, which have been used as positional cloning resources as well as to support directed and genome-wide sequencing efforts. This unit describes a robust, large-scale procedure for generation of agarose gel–based clone fingerprints from BAC clones. Keywords: Restriction Digest fingerprint; bacterial artificial chromosome (BAC); BAC DNA isolation; agarose gel electrophoresis; BAC fingerprint map
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Large-scale BAC clone Restriction Digest fingerprinting.
Current protocols in human genetics, 2007Co-Authors: Carrie A Mathewson, Jacqueline E Schein, Marco A MarraAbstract:Restriction Digest fingerprinting is a common method for characterizing large insert genomic clones, e.g., bacterial artificial chromosome (BAC), P1 artificial chromosome (PAC) and Fosmid clones. This clone fingerprinting method has been widely applied in the construction of clone-based physical maps, which have been used as positional cloning resources as well as to support directed and genome-wide sequencing efforts. This unit describes a robust, large-scale procedure for generation of agarose gel-based clone fingerprints from BAC clones.
