scholarly journals A physical map of the human regulator of complement activation gene cluster linking the complement genes CR1, CR2, DAF, and C4BP.

1988 ◽  
Vol 167 (2) ◽  
pp. 664-669 ◽  
Author(s):  
J Rey-Campos ◽  
P Rubinstein ◽  
S Rodriguez de Cordoba
Keyword(s):  
Gene Cluster ◽  
Complement Activation ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Pulsed Field Gel Electrophoresis ◽  
Complement Components ◽  
Pulsed Field ◽  
Human Genes ◽  
Tight Linkage ◽  
Genes Encoding

We report the organization of the human genes encoding the complement components C4-binding protein (C4BP), C3b/C4b receptor (CR1), decay accelerating factor (DAF), and C3dg receptor (CR2) within the regulator of complement activation (RCA) gene cluster. Using pulsed field gel electrophoresis analysis these genes have been physically linked and aligned as CR1-CR2-DAF-C4BP in an 800-kb DNA segment. The very tight linkage between the CR1 and the C4BP loci, contrasted with the relative long DNA distance between these genes, suggests the existence of mechanisms interfering with recombination within the RCA gene cluster.

Construction of a genomic map ofMoraxella (Branhamella) catarrhalisATCC 25238 and physical mapping of virulence-associated genes

1999 ◽  
Vol 45 (4) ◽  
pp. 299-303 ◽  
Author(s):  
K T Nguyen ◽  
E J Hansen ◽  
M A Farinha
Keyword(s):  
Gel Electrophoresis ◽  
Moraxella Catarrhalis ◽  
Southern Hybridization ◽  
Physical Map ◽  
Outer Membrane Proteins ◽  
Restriction Enzymes ◽  
Pulsed Field Gel Electrophoresis ◽  
Respiratory Pathogens ◽  
Circular Chromosome ◽  
Pulsed Field

A physical genome map of the Moraxella catarrhalis type strain (ATCC 25238) has been constructed using pulsed field gel electrophoresis. Macrorestriction analyses of the genome of M. catarrhalis were performed by digestion with the restriction enzymes SmaI, NotI, and RsrII, which cleave the single circular chromosome into 9, 10, and 6 fragments, respectively. The chromosomal fragments generated by pulsed field gel electrophoresis were converted to a linkage map utilizing a combination of partial digestions, and cross-hybridizations. Moraxella catarrhalis, like a number of other respiratory pathogens, has a relatively small genome estimated at 1750 kilobase pairs or about 40% of the size of the Escherichia coli genome. The locations of the four ribosomal RNA operons (rrnLS) were determined by Southern hybridization and by digestion with I-CeuI endonuclease. A number of genes involved in virulence have been placed onto the physical map by Southern hybridization including those encoding the predominant outer-membrane proteins and the chromosomal gene encoding beta-lactamase.Key words: Moraxella catarrhalis, physical map, genome analysis, pulsed-field gel electrophoresis, virulence.

scholarly journals Plasmid-Borne Type E Neurotoxin Gene Clusters in Clostridium botulinum Strains

2013 ◽  
Vol 79 (12) ◽  
pp. 3856-3859 ◽  
Author(s):  
Zhen Zhang ◽  
Hannamari Hintsa ◽  
Ying Chen ◽  
Hannu Korkeala ◽  
Miia Lindström
Keyword(s):  
Gene Cluster ◽  
Gel Electrophoresis ◽  
Clostridium Botulinum ◽  
Southern Hybridization ◽  
Gene Clusters ◽  
Pulsed Field Gel Electrophoresis ◽  
Content Type ◽  
Pulsed Field ◽  
Large Plasmids

ABSTRACTA collection of 36Clostridium botulinumtype E strains was examined by pulsed-field gel electrophoresis (PFGE) and Southern hybridization with probes targeted tobotEandorfX1in the neurotoxin gene cluster. Three strains were found to contain neurotoxin subtype E1 gene clusters in large plasmids of about 146 kb in size.

Staphylococcal Poisoning Foodborne Outbreak: Epidemiological Investigation and Strain Genotyping

2013 ◽  
Vol 76 (12) ◽  
pp. 2093-2098 ◽  
Author(s):  
S. GALLINA ◽  
D. M. BIANCHI ◽  
A. BELLIO ◽  
C. NOGAROL ◽  
G. MACORI ◽  
...  
Keyword(s):  
Nasal Mucosa ◽  
Biological Samples ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Food Handler ◽  
Staphylococcal Protein A ◽  
Spa Typing ◽  
Pulsed Field ◽  
Genes Encoding ◽  
Genetic Profiles

In June 2011, an outbreak of Staphylococcus aureus enterotoxin food poisoning gastroenteritis occurred in Turin, Italy, following a catered dinner party at a private home. Within a few hours, 26 of the 47 guests experienced gastrointestinal illness, and 9 were hospitalized. A retrospective cohort study using a standardized questionnaire was carried out, and the risk ratios for each food item were calculated. The analysis indicated consumption of seafood salad as the most probable cause of the outbreak (risk ratio = 11.72; 95% confidence interval, 1.75 to 78.54). Biological samples were collected from four of the hospitalized guests (stool and vomit), nasal mucosa swabs from three food handlers employed with the caterer, and available food residuals. All stool and vomit samples tested positive for enterotoxigenic S. aureus. As residues of the seafood salad were no longer available for sampling, suspected contamination could not be verified. However, no other food was found contaminated by S. aureus or its enterotoxins. All isolates from the biological samples were characterized at the genomic level by means of two multiplex PCR protocols to determine the presence of genes encoding staphylococcal enterotoxins, pulsed-field gel electrophoresis and staphylococcal protein A gene (spa) typing to describe their genetic profiles. All the isolates presented genes encoding SEA and SEI; the pulsed-field gel electrophoresis genetic profiles revealed the same pulsotype in the microorganism isolated from the hospitalized guests as in one of the isolates from a food handler's nasal mucosa, and the spa-typing analysis reported two closely related spa types (t701 and t267), implicating the food handler as the most likely outbreak source.

scholarly journals Monitoring and Characterization of Extended-Spectrum β-Lactamases in Escherichia coli Strains from Healthy and Sick Animals in Spain in 2003

2005 ◽  
Vol 49 (3) ◽  
pp. 1262-1264 ◽  
Author(s):  
Laura Briñas ◽  
Miguel Angel Moreno ◽  
Tirushet Teshager ◽  
Yolanda Sáenz ◽  
María Concepción Porrero ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Extended Spectrum ◽  
Genes Encoding ◽  
M 14

ABSTRACT Genes encoding CTX-M-14, CTX-M-9, CTX-M-1, CTX-M-32, SHV-12, TEM-52, or CMY-2 β-lactamases were detected in 21 Escherichia coli strains recovered during 2003 from sick animals (11 of 459 [2.4%] strains) and healthy animals (10 of 158 [6.3%] strains) in Spain. Twelve of these strains harbored bla CTX-M genes and showed unrelated pulsed-field gel electrophoresis patterns.

scholarly journals Comparison of the beta-lactamase gene cluster in clonally distinct strains of Enterococcus faecalis.

1996 ◽  
Vol 40 (5) ◽  
pp. 1170-1174 ◽  
Author(s):  
J F Tomayko ◽  
K K Zscheck ◽  
K V Singh ◽  
B E Murray
Keyword(s):  
Gene Cluster ◽  
Enterococcus Faecalis ◽  
Gel Electrophoresis ◽  
Considerable Change ◽  
Pulsed Field Gel Electrophoresis ◽  
Enzyme Analysis ◽  
Beta Lactamase ◽  
Pulsed Field ◽  
Large Clone ◽  
Lactamase Gene

Ten beta-lactamase-producing Enterococcus faecalis isolates were examined for the presence of the staphylococcal beta-lactamase repressor and antirepressor genes. Four isolates, previously shown to be unrelated to each other by pulsed-field gel electrophoresis analysis, were positive for both genes by PCR, although beta-lactamase production was not induced with methicillin. Six isolates, previously shown to be clonally related, were negative for both genes by PCR. The blaZ sequences of eight beta-lactamase-producing E. faecalis isolates were determined. Seven isolates from five distinct clones had sequences identical to that previously reported for E. faecalis HH22, regardless of whether the repressor or antirepressor was demonstrated by PCR. However, blaZ from one isolate differed from those of the other enterococci by 11 nucleotides; this isolate is part of the large clone, as defined by pulsed-field gel electrophoresis and multilocus enzyme analysis, that includes HH22. These findings suggest either that enterococci have acquired the bla gene cluster from more than one source or that the gene cluster has undergone considerable change since acquisition by this clone.

A bacterial artificial chromosome based physical map of the Ustilago maydis genome

Genome ◽  
2005 ◽  
Vol 48 (2) ◽  
pp. 207-216 ◽  
Author(s):  
Khalid Meksem ◽  
Jeffry Shultz ◽  
Faiza Tebbji ◽  
Aziz Jamai ◽  
Jürgen Henrich ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Artificial Chromosome ◽  
Ustilago Maydis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Artificial Chromosomes ◽  
Bac Clones ◽  
Copy Dna

Ustilago maydis, a basidiomycete, is a model organism among phytopathogenic fungi. A physical map of U. maydis strain 521 was developed from bacterial artificial chromosome (BAC) clones. BAC fingerprints used polyacrylamide gel electrophoresis to separate restriction fragments. Fragments were labeled at the HindIII site and codigested with HaeIII to reduce fragments to 50–750 bp. Contiguous overlapping sets of clones (contigs) were assembled at nine stringencies (from P ≤ 1 x 10–6 to 1 x 10–24). Each assembly nucleated contigs with different percentages of bands overlapping between clones (from 20% to 97%). The number of clones per contig decreased linearly from 41 to 12 from P ≤ 1 x 10–7 to 1 x 10–12. The number of separate contigs increased from 56 to 150 over the same range. A hybridization-based physical map of the same BAC clones was compared with the fingerprint contigs built at P ≤ 1 × 10–7. The two methods provided consistent physical maps that were largely validated by genome sequence. The combined hybridization and fingerprint physical map provided a minimum tile path composed of 258 BAC clones (18–20 Mbp) distributed among 28 merged contigs. The genome of U. maydis was estimated to be 20.5 Mbp by pulsed-field gel electrophoresis and 24 Mbp by BAC fingerprints. There were 23 separate chromosomes inferred by both pulsed-field gel electrophoresis and fingerprint contigs. Only 11 of the tile path BAC clones contained recognizable centromere, telomere, and subtelomere repeats (high-copy DNA), suggesting that repeats caused some false merges. There were 247 tile path BAC clones that encompassed about 17.5 Mbp of low-copy DNA sequence. BAC clones are available for repeat and unique gene cluster analysis including tDNA-mediated transformation. Program FingerPrint Contigs maps aligned with each chromosome can be viewed at http://www.siu.edu/~meksem/ustilago_maydis/.Key words: Ustilago maydis, physical map, bacterial artificial chromosomes, whole-genome sequencing.

scholarly journals Mapping of Escherichia coli chromosomal Tn5 and F insertions by pulsed field gel electrophoresis.

Genetics ◽  
1988 ◽  
Vol 119 (2) ◽  
pp. 227-236
Author(s):  
C L Smith ◽  
R D Kolodner
Keyword(s):  
Escherichia Coli ◽  
Physical Mapping ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Pulsed Field Gel Electrophoresis ◽  
Genetic Maps ◽  
Low Resolution ◽  
Transposon Tn5 ◽  
Pulsed Field ◽  
E Coli

Abstract A low resolution Not I physical map of Escherichia coli was recently constructed. In this report we demonstrated that this map can be used to map Tn5 and F insertions physically. The transposon, Tn5, contains Not I recognition sequences in its IS50 sequences. F plasmid contains an unmapped Not I site. Hence, the location of Tn5 and F in the chromosome can be mapped by identifying the location of the introduced Not I sites using pulsed field gel electrophoresis. The physical mapping of genetically mapped Tn5 insertions confirm the previously constructed Not I map and helps align the E. coli physical and genetic maps. The use of Tn5 can assist the construction of both physical and genetic maps for microorganisms lacking such maps. Variations on this approach will facilitate physical mapping with a wide variety of organisms, enzymes, and genetic elements.

scholarly journals Serotype 19F Multiresistant Pneumococcal Clone Harboring Two Erythromycin Resistance Determinants [erm(B) and mef(A)] in South Africa

2001 ◽  
Vol 45 (5) ◽  
pp. 1595-1598 ◽  
Author(s):  
Lesley McGee ◽  
Keith P. Klugman ◽  
Avril Wasas ◽  
Thora Capper ◽  
Adrian Brink
Keyword(s):  
South Africa ◽  
Streptococcus Pneumoniae ◽  
Private Sector ◽  
Dna Fingerprinting ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Erythromycin Resistance ◽  
Pulsed Field ◽  
Resistance Determinants ◽  
Genes Encoding

ABSTRACT One hundred eighteen erythromycin-resistant Streptococcus pneumoniae (ERSP) strains (MICs of ≥0.5 μg/ml) from five laboratories serving the private sector in South Africa were analyzed for the genes encoding resistance to macrolides. Sixty-seven ERSP strains (56.8%) contained the erm(B) gene, and 15 isolates (12.7%) contained the mef(A) gene. Thirty-six isolates (30.5%) harbored both the erm(B) and mef(A) genes and were highly resistant to erythromycin and clindamycin. DNA fingerprinting by BOX-PCR and pulsed-field gel electrophoresis identified 83% of these strains as belonging to a single multiresistant serotype 19F clone.

Mapping of genes encoding glycoside hydrolases on the chromosome of Cellulomonas fimi

2001 ◽  
Vol 47 (12) ◽  
pp. 1063-1067 ◽  
Author(s):  
Dominik Stoll
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Glycoside Hydrolases ◽  
Circular Chromosome ◽  
Cellulomonas Fimi ◽  
Pulsed Field ◽  
A Genome ◽  
Genome Map ◽  
Genes Encoding ◽  
Single Circular Chromosome

Cellulomonas fimi genomic DNA was digested with HpaI, MunI, HindIII, and NsiI, producing fragments ranging in size from 20 to 1400 kbp that were resolved by pulsed field gel electrophoresis. Genetic and physical linkages were determined by Southern blotting and were used to construct a genome map. Cellulomonas fimi has a single circular chromosome of approx. 4000 kbp. Except for two closely linked genes, cbh6A and cel5A, the genes known to encode glycoside hydrolases are scattered widely on the chromosome.Key words: Cellulomonas fimi, genome map, pulsed field gel electrophoresis, glycoside hydrolases.

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