Persistence of Enterobacteriaceae in female adults of the biting gnat Culicoides variipennis (Diptera: Ceratopogonidae).
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The selC gene from E. coli codes for a tRNA species (tRNA(UCASer] which is aminoacylated with L-serine and which cotranslationally inserts selenocysteine into selenoproteins. By means of Southern hybridization it was demonstrated that this gene occurs in all enterobacteria tested. To assess whether the unique primary and secondary structural features of the E. coli selC gene product are conserved in that of other organisms, the selC homologue from Proteus vulgaris was cloned and sequenced. It was found that the Proteus selC gene differs from the E. coli counterpart in only six nucleotides, that it displays the same unique properties and that it is expressed and functions in E. coli. This indicates that the unique mechanism of selenocysteine incorporation is not restricted to E. coli but has been conserved as a uniform biochemical process.
Tryptophan biosynthesis in Escherichia coli is regulated by the product of the trpR gene, the tryptophan (Trp) repressor. Trp aporepressor binds the corepressor, L-tryptophan, to form a holorepressor complex, which binds trp operator DNA tightly, and inhibits transcription of the tryptophan biosynthetic operon. The conservation of trp operator sequences among enteric Gram-negative bacteria suggests that trpR genes from other bacterial species can be cloned by complementation in E. coli. To clone trpR homologues, a deletion of the E. coli trpR gene, delta trpR504, was made on a plasmid by site-directed mutagenesis, then crossed onto the E. coli genome. Plasmid clones of the trpR genes of Enterobacter aerogenes and Enterobacter cloacae were isolated by complementation of the delta trpR504 allele, scored as the ability to repress beta-galactosidase synthesis from a prophage-borne trpE-lacZ gene fusion. The predicted amino acid sequences of four enteric TrpR proteins show differences, clustered on the backside of the folded repressor, opposite the DNA-binding helix-turn-helix substructures. These differences are predicted to have little effect on the interactions of the aporepressor with tryptophan, holorepressor with operator DNA, or tandemly bound holorepressor dimers with one another. Although there is some variation observed at the dimer interface, interactions predicted to stabilize the interface are conserved. The phylogenetic relationships revealed by the TrpR amino acid sequence alignment agree with the results of others.
We describe EnteriX, a suite of three web-based visualization tools for graphically portraying alignment information from comparisons among several fixed and user-supplied sequences from related enterobacterial species, anchored on a reference genome (http://bio.cse.psu.edu/). The first visualization, Enteric, displays stacked pairwise alignments between a reference genome and each of the related bacteria, represented schematically as PIPs (Percent Identity Plots). Encoded in the views are large-scale genomic rearrangement events and functional landmarks. The second visualization, Menteric, computes and displays 1 Kb views of nucleotide-level multiple alignments of the sequences, together with annotations of genes, regulatory sites and conserved regions. The third, a Java-based tool named Maj, displays alignment information in two formats, corresponding roughly to the Enteric and Menteric views, and adds zoom-in capabilities. The uses of such tools are diverse, from examining the multiple sequence alignment to infer conserved sites with potential regulatory roles, to scrutinizing the commonalities and differences between the genomes for pathogenicity or phylogenetic studies. The EnteriX suite currently includes >15 enterobacterial genomes, generates views centered on four different anchor genomes and provides support for including user sequences in the alignments.
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Vaccines prepared from Gram-negative bacteria isolated from the stools of HLA-B27 positive AS patients were used to immunize rabbits. Three of the sera obtained were lytic in vitro for the mononuclear cells of HLA-B27 positive AS patients. One of these sera discriminated between AS patients and healthy HLA-B27 positive individuals. Cytolysis was determined in an automated, non-radioactive assay based on the release of carboxyfluorescein diacetate and the incorporation of propidium iodide.
Sera from patients infected with Escherichia coli, Proteus, Klebsiella, and Serratia were studied for precipitins to ultrasonic extracts of these organisms in gel-diffusion plates. Sera from 66 per cent of these patients contained precipitins when initially tested. Twenty-four per cent of sera tested in the first week after onset of infection contained precipitins, but this rose to 78 per cent by the third week. Cross-reactions of sera with Pseudomonas antigens were unusual, but were common with other enterobacterial antigen extracts. However, higher titers were usually present to homologous as compared to heterologus antigens. Sera from seven patients contained precipitins to a common enterobacterial antigen. Precipitins to E. coli, Proteus, Klebsiella, and Serratia were detected in only a small proportion of control sera.
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Bacterial strains isolated from wide ranges of nematode hosts and geographic sources and strains isolated from human clinical specimens were used to assess the taxonomic structure of the genus Photorhabdus. The following two methods were used: DNA relatedness and phenotypic characterization. Analysis of the DNA relatedness data revealed that all of the strains studied were congeneric and that the genus Photorhabdus is, on the basis of DNA relatedness data, more homogeneous than the other genus of nematode-symbiotic bacteria, the genus Xenorhabdus. In contrast to previous reports, only two DNA relatedness groups were identified in the genus Photorhabdus. These groups corresponded to the symbiotic strains and the clinical strains. There appeared to be some subgroups within the symbiotic strain group on the basis of the interactions of the strains with nematodes, which corresponded to some extent with the DNA relatedness data. However, there were significant ambiguities in the DNA relatedness data, and this group could not be subdivided on the basis of DNA relatedness data or phenotypic data. The distinct functional differences within and between the DNA relatedness groups of symbiotic Photorhabdus strains indicated that there are biologically significant sub-groups within the genus Photorhabdus that cannot be defined at this time. Further investigation of the taxonomy of Photorhabdus by using different approaches and a suitably wide range of strains is recommended. However, it is clear that the clinical strains form a recognizable subgroup within the genus even though no formal subtaxon can be defined at this time.
Data from a study of both phases of 21 strains of Xenorhabdus examined for 240 characters were subjected to numerical analysis. Only 60 characters were used for the analyses, since 169 characters were common to all isolates, and the acidification data essentially duplicated the assimilation tests. The data were arranged in seven ways to determine the significance of characters affected by phase change and of weak responses. Most of the analyses involved calculation of similarities by the Jaccard coefficient and clustering by single linkage, complete linkage and centroid sorting algorithms. The resultant dendrograms emphasized the importance of recognizing phase-related characteristics in examining the taxonomy of Xenorhabdus. They also demonstrated a close correspondence between the taxonomic groupings of Xenorhabdus and those of their nematode associates. It is proposed that the subspecies of X. nematophilus be elevated to species, X. nematophilus, X. bovienii, X. poinarii and X. beddingii.
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