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  • 1
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Multiple antibiotic-resistant Shigella dysenteriae type 1 isolates from a recent epidemic in West Bengal (India) showed identical plasmid patterns. All isolates were resistant to ampicillin (Am), chloramphenicol (Cm), tetracycline (Tc), streptomycin (Sm) and trimethoprim (Tp) and contained 6 plasmids, ranging from 2.5–120 kb. The Am resistance determinant was located on the 120 kb plasmid. This plasmid was unstable when the S. dysenteriae strains were grown above 37°C. The Bangladesh strains of S. dysenteriae type 1 showed identical plasmid patterns, except that many isolates were Am-sensitive and lacked the 120 kb plasmid. In strains from both Bangladesh and West Bengal, predominantly group-B plasmids conferred resistance to Cm and Tc. Comparisons of EcoR1 fragments generated from the total plasmid DNA content of each strain support the view that the plasmids present in the S. dysenteriae type 1 strains isolated from all recent epidemics in India and Bangladesh were identical.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 84 (1991), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Growth conditions play a major role in expression of virulence by Shigella spp. both in vitro (adherence and internalization in eukaryotic host cells) and in vivo (keratoconjunctivitis). Optimized expression of virulence required anaerobic growth to log phase in particular media such as brain heart infusion broth. Kinetic studies of guinea pig eye infections showed that as few as 2 × 105S. dysenteriae CG097 or S. flexneri M90T, grown under these optimized conditions, produced keratoconjuctivitis in 15 h. In vitro studies demonstrated that adherence to and invasion of Henle 407 cells, at 37°C, by organisms grown under these optimized conditions, were significantly greater than when organisms were grown aerobically under the same conditions.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 55 (1988), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Outer membranes of Shigella species and E. coli K-12 carrying large invasive plasmids and isogenic non-invasive strains without plasmids were analyzed by SDS-PAGE. The immunoblotting analysis of the outer membrane proteins of these bacteria was performed with monoclonal antibody (mAb) made against A and B subunits of Shiga-like toxin (SLT). The SLT was detected in the outer membranes of S. dysenteriae 1 IDBM11, S. sonnei PNS20, S. flexneri M90T, S. dysenteriae 60R, and E. coli K-12 strain AB2463. The two other E. coli K-12 strains, C600 and 933J were included as controls for low and high toxin producers respectively. The outer membrane protein band of molecular weight 70 kDa was common to all bacterial strains studied. The most prominent band of 70 kDa protein was seen to be present in the high toxin producing plasmidless strain of S. dysenteriae 60R and the lysogenic strain of E. coli 933J. The invasive strains of S. dysenteriae 1 and S. flexneri M90T which carry the large invasive plasmids showed the least prominent band of 70 kDa protein.The immunoblotting analysis of Shiga-toxin partially purified from the S. dysenteriae 60R strain revealed the absence of 70 kDa band on SDS-PAGE, instead the two dissociated subunits were seen. Furthermore, periplasmic Shiga-toxin proteins also showed the complete dissociation into A and B subunits. However, under the same denaturing conditions, the 70 kDa protein band cross-reacting with mAb against A and B subunits was still present in the outer membranes of all different strains.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 37 (1986), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract A genetic determinant for serum resistance, designated iss, has been found previously on the colicinogenic plasmid ColV2-K94. In this work we have identified a second serum resistance gene, traT, on ColV2-K94. The serum resistance mediated by derivatives of ColV2-K94 was due to presence of one or both of the iss and traT genes. Plasmid pWS12 (TraT+ Iss+) contained the kanamycin (Km) resistance transposon Tn903 inserted near the origin of replication of ColV2-K94, and plasmids pWS15 (TraT+), pWS16 (TraT+) and pWS18 (TraT+ Iss+) were deletion derivatives of pWS12 constructed in vitro and in vivo. pWS12 and pWS18 conferred a 20-fold increase in relative resistance to 20% guinea pig serum when introduced into the serum-susceptible, genetically defined recA strain of Escherichia coli K-12, AB2463. Plasmids pWS15 and pWS16, from which iss had been deleted, still conferred 5-fold increases in relative resistance on AB2463. The level of resistance conferred on this strain by the antibiotic resistance plasmid R100–1 (which expresses the traT serum resistance gene) was comparable to that of plasmids pWS15 and pWS16. The 25-kDa traT gene product was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the outer membrane proteins of strain AB2463 carrying ColV2-K94. This protein cross-reacted immunologically with the traT protein expressed by F or R100–1. Our results indicated that both traT and iss are capable of mediating serum resistance in ColV2-K94.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Four enteropathogenic Escherichia coli strains were studied with respect to their antibiotic resistance characters, plasmid patterns, toxin production and haemagglutination properties. Two of these strains showed multiple antibiotic resistance characters, although all possessed several plasmids of varying sizes. One of the strains DD-41 showed the presence of a non-fimbrial cell-associated mannose-resistant haemagglutinin (MRHA) which was encoded by a 70 MDa plasmid. Conjugation experiments demonstrated that this MRHA-containing plasmid also coded for ampicillin and tetracycline resistance factors and was self-transmissible.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 83 (1991), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract: Evidence is presented that a high level Shiga toxin-producing strain Shigella dysenteriae 60R adheres to and invades the epithelial cell lines Hct8 and Henle 407. The invasive phenotype of S. dysenteriae 60R differs in four ways from the heretofore studied invasive Shigella phenotypes. First, S. dysenteriae 60R lacks the virulence plasmid characteristic of other invasive Shigella spp. and enteroinvasive Escherichia coli. Second, hybridization studies show that the known ipa genes are neither present in the chromosome nor in the small plasmid of 60R. Third, 60R adheres to and invades Hct8 and Henle 407 cells at 37°C as well as at 30°C. Fourth, the phenotype of adherence and invasion of 60R is remarkably stable, even during prolonged growth in laboratory media and storage.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 146 (1976), S. 215-231 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fused F prime factor was obtained from a mating of arecA donor carrying an F' factor containing the genesmetBJF, ppc andargECBH (KLF5) with arecA recipient carrying an F' factor containingatt80, trp andlac (F155). Lysogenization of this fused F-prime factor with λcI857 hϕ80 phage followed by thermoinduction produced the transducing phages ϕ80dmetBJF and ϕ80dppcargECBH. This kind of fusion provides a general procedure for the construction of transducing phages carrying genes from different regions of theE. coli genome. To understand the mechanism of this fusion, the parental F prime factors (F155 and KLF5) were analyzed by the electron microscope heteroduplex technique. F155 has a length of 176±3 kilobases including two substitutions. The F sequence 0 F-2.8 F has been substituted by 53 kb of chromosomal DNA including thelac operon and the F sequences 8.5 F-16.3 F has been substituted by 27 kb of a chromosomal sequence includingatt80 and thetrp operon. KLF5 contains 221±4 kilobases of DNA (molecular weight, 148 megadaltons). It contains complete F and the segment of theE. coli chromosome frompolA torif. The F sequence 2.8 F-8.5 F known to be involved in F specific recombination inrecA + andRecA backgrounds occurs twice on KLF5, once at each of the junctions of F DNA with chromosomal DNA. The population of closed circular plasmid molecules extracted from KLF5-containing strains is heterogeneous. It is proposed that this heterogeneity is due to intramolecular recombination events occurring in KLF5 between the duplicated 2.8 F-8.5 F sequences. Such recombination can account for the genetic instability of KLF5 observed in bothrecA + andrecA hosts. The F sequence 2.8 F-8.5 F (also called γδ) is one of the characterized integration sequences on F. A model for the fusion of the parental F prime factors is proposed in which recombination between γδ sequences bringsatt80 close to themetBJF genes. This is followed by a deletion of an F'lac factor. The resulting fused F' factor still carries two γδ sequences and is therefore expected to be unstable. The closed circular molecules isolated from the fused F' containing strains show two different sizes of molecules. Genetic and physical analyses of these molecules are in agreement with the predicted instability of the fused F' factor and the existance of the γδ sequence in the ϕ80dmet phages isolated from fused F' and previously analyzed by the electron microscope heteroduplex technique.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 193 (1984), S. 543-553 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Four E. coli Hfr strains, representing stable (Hfr Cavalli), moderately stable (AB312) and unstable (Ra-1, Ra-2) Hfr states, were used in the isolation of a series of F′ plasmids. Type II F′s were found to be the most prevalent F′ plasmid formed from all of the Hfrs, while the percentages of Δtra F′s increased as the stability of the Hfr increased. Two observations suggested that F′ formation in unstable Hfrs like Ra-2 may proceed through a type II F′ precursor. First, the major F′ products of Ra-2 are tra + type II F′s and, second, other F′ types (I, II) and classes (tra +, Δtra) from Ra-2 appeared to be deletion derivatives of a larger F′ progenitor. By monitoring the molecular changes that occur when the Ra-2 derived type II F′ pWS200 is transferred from one recA host to another, we have found that all F′ types and classes can be generated from pWS200 in a recA-independent manner. F sequences involved in the genetic conversions of pWS200 include the oriT locus and the directly repeated γδ junctions of F and chromosomal DNA. A model for the formation of F′s in unstable Hfrs is postulated in which a tra + type II F′ primary excision product is seen to be modified, through recA-independent processes, to other F′ types and classes. This model differs from the current model of F′ formation in that independent excision events from the Hfr chromosome are not seen as the source of type I and type II F′s. These studies have also shown that the formation of δtra F′s is a recA-independent process that can occur from the F′ and Hfr states, that γδ-mediated deletions in pWS200 often demonstrate regional specificity in having endpoints near the ilv operon and that genetic alterations in either replication origin of pWS200 (F oriV, chromosomal oriC) stabilize the replication of this “mini-Hfr” cointegrate.
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  • 9
    ISSN: 1617-4623
    Keywords: Bacteriophage lambda ; IS2 ; Serum survival ; ColV2-K94
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Escherichia coli K12 cells carrying a cloned 1.4 kb HindIII fragment from plasmid ColV2-K94, showed increased survival in guinea pig serum. The recombinant plasmid also conferred group II surface exclusion, i.e. the cells were reduced in recipient ability towards the incoming plasmid R538drd in conjugation experiments. Southern blotting suggested homology with bacteriophage lambda DNA and to the insertion element IS2. Determination of the DNA sequence of the fragment demonstrated the presence of a truncated IS2 (165 bp), separated by 250 bp from a 900 bp stretch of homology with lambda DNA, beginning within the Rz gene and continuing in the rightward direction on the lambda map. A 97 amino acid open reading frame (ORF) adjacent to Rz and on the opposite strand, remained intact in iss, with several amino acid changes. The ORF in iss is preceded by sequences resembling prokaryotic ribosome binding sites and promoters.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transducing λ bacteriophages have been isolated which carry the divergently transcribed argECBH operon of E. coli K12 and various portions of the adjacent ppc and bfe chromosomal regions. They were recovered from lysates prepared by the procedure of Schrenk and Weisberg using a Ppc+ Arg+ Bfe+ strain carrying a deletion of the usual attachment site of λ. Heteroduplex DNA mapping of these λdarg and of the ϕ80 darg isolated by B. Konrad indicates that the two kinds of phages carry the arg cluster in opposite orientations, a situation favorable for the isolation of argECBH DNA. A physical map of the ppc argECBH bfe region including 2 unusual attachment sites of λ has been constructed. The localization of the end points of certain arg deletions provides a useful reference framework for the currently pursued mapping of mutations affecting the control of divergent transcription and for the location of restriction enzyme cleavage sites in the arg region.
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