Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Twenty-five isolates of the bacterium Pasteurella multocida were characterized (fingerprinted) phenotypically and genotypically in order to compare the abilities of various techniques to differentiate strains for epidemiologic studies of fowl cholera. Isolates were obtained over a 16-month period from turkeys dying from fowl cholera (six outbreak flocks) and from wildlife captured on premises with a history of the disease. The characteristics compared included (i) serotype, (ii) subspecies, (iii) antibiogram, (iv) presence of plasmid DNA, (v) restriction endonuclease analysis patterns of whole-cell DNA, and (vi) ribotype. Ribotyping, a method of highlighting DNA restriction site heterogeneity by using an rRNA probe, worked well for differentiating the strains of P. multocida when the majority of the other techniques could not. Ribotyping results correlated directly with and confirmed results obtained from restriction endonuclease analysis. Ribotyping demonstrated the presence of up to three strains of P. multocida in one outbreak flock, recurrence of a single strain in five of the flocks over an 11-month period, and the presence of common strains in turkeys and wildlife on the premises.
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PMID:Use of an rRNA probe and restriction endonuclease analysis to fingerprint Pasteurella multocida isolated from turkeys and wildlife. 276 71

Operon fusions for the Escherichia coli heat-labile enterotoxin type IIa (LT-IIa) operon were isolated and characterized. The LT-IIa genes are organized in a transcriptional unit similar to those of cholera toxin (CT) and the closely related E. coli heat-labile toxin type I (LT-I, with subtypes LTh-I and LTp-I). The nucleotide sequence of the LT-IIa genes was determined and compared with the sequences of LTh-I and CT. The A subunit gene of LT-IIa was found to be 57% homologous with the A subunit gene of LTh-I and 55% homologous with the A gene of CT. Most of the homology derived from the region of the A gene which encodes the A1 fragment. The B gene of LT-IIa was not homologous with the B gene of LTh-I or CT. DNA probes containing various portions of the LT-IIa genes and adjacent sequences were used for hybridization studies with restriction endonuclease fragments of DNA from a collection of LT-II-producing strains. These studies showed that a probe containing much of the A subunit gene hybridized well to DNA from the various strains, but a probe for the B subunit gene did not.
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PMID:Genetics of type IIa heat-labile enterotoxin of Escherichia coli: operon fusions, nucleotide sequence, and hybridization studies. 282 67

The P plasmid of Vibrio cholerae is a derepressed sex factor restricted to V. cholerae and has been shown to express surface exclusion. We have isolated the plasmids of strain V58 and have found that in addition to P, two further cryptic plasmids are also present. P has a size of 68 kb as determined by both electron microscopy and restriction endonuclease analysis. These other plasmids are 34 and 4.7 kb in size. Restriction maps of P and the larger cryptic plasmid have been determined. It has been demonstrated that P differs from the standard Inc group test plasmids and also expresses a surface exclusion system. The ability of the type Inc plasmids to be transferred to V. cholerae by either liquid or filter matings and the stability of these plasmids in V. cholerae have also been examined.
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PMID:Characterization and restriction analysis of the P sex factor and the cryptic plasmid of Vibrio cholerae strain V58. 282 2

A cosmid gene bank of Vibrio cholerae 395, classical Ogawa, was screened in Escherichia coli HB101 for expression of the vibrio neuraminidase (NANase) gene nanH (N-acylneuraminate glycohydrolase). Positive clones were identified by their ability to cleave the fluorogenic NANase substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid. Seven NANase-positive clones were detected after screening 683 cosmid isolates with a rapid, qualitative plate assay method. The nanH gene was subcloned from one of the cosmids and was located within a 4.8-kilobase-pair BglII restriction endonuclease fragment. Evidence that nanH was the NANase structural gene was obtained by transposon mutagenesis and by purification and comparison of the cloned gene product with the secreted NANase purified from the parent V. cholerae strain. The sequence of the first 20 amino-terminal amino acids of the secreted NANase purified from V. cholerae was determined by automated Edman degradation and matched perfectly with the amino acid sequence predicted from nucleotide sequencing of nanH. The sequence data also revealed the existence of a potential signal peptide that was apparently processed from NANase in both V. cholerae and E. coli. In contrast to V. cholerae, E. coli nanH+ clones did not secrete NANase into the growth medium, retaining most of the enzyme in the periplasmic compartment. Kinetic studies in V. cholerae showed that nanH expression and NANase secretion were temporally correlated as cells in batch culture entered late-exponential-phase growth. Similar kinetics were observed in at least one of the E. coli nanH+ clones, suggesting that nanH expression in E. coli might be controlled by some of the same signals as in the parent V. cholerae strain.
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PMID:Cloning and expression of the Vibrio cholerae neuraminidase gene nanH in Escherichia coli. 283 65

Plasmid profile analysis, plasmid and chromosomal bacterial restriction endonuclease DNA analysis, and DNA hybridization are increasingly used in clinical microbiology and epidemiology. These techniques have been applied, singly and in combination, to investigations of outbreaks, including those of diarrheal diseases caused by pandemic Vibrio cholerae, enterotoxigenic and enterohemorrhagic Escherichia coli, Salmonella muenchen, and Salmonella typhimurium. The techniques have been critical in the unraveling of outbreaks of disease caused by nosocomial and community-acquired pathogens. Although there are limitations to these methods, their applications have important ramifications for basic science and public health.
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PMID:Molecular epidemiology of bacterial infections: examples of methodology and of investigations of outbreaks. 302 88

The structure of the cholera toxin operon and the location of A and B toxin subunits have been studied by the Southern blot hybridization on filters. The gene coding for the synthesis of the cholera toxin B-subunit has been cloned in the vector plasmid pBR322. The structural gene of A-subunit has been partially deleted by the restriction endonuclease Bal31 digestion. The size of the 250 b. p. deletion has been defined by electron microscopy. The production of the cholera toxin B-subunit in Escherichia coli K12 cells has been studied.
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PMID:[Construction of recombinant plasmids encoding the biosynthesis of the beta-subunit of cholera toxin]. 303 59

Restriction endonuclease-generated polynucleotide and synthetically produced oligonucleotide gene probes used in colony hybridization assays proved to be efficient for the detection and differentiation of enterotoxigenic Escherichia coli. To compare their relative efficiencies, these two sets of probes were radiolabeled with 32P and were applied to 74 strains of E. coli with known enterotoxin profiles and to 156 previously unexamined E. coli isolates. The enterotoxigenic bacteria Vibrio cholerae O1, Vibrio cholerae non-O1 (NAG), Yersinia enterocolitica, and E. coli harboring the plasmid vectors of the polynucleotide gene probes were examined for further evaluation of probe specificity. The two classes of probes showed a perfect concordance in their specific detection and differentiation of enterotoxigenic E. coli. In the analysis of six strains, the signal strength on autoradiography after hybridization with oligonucleotides was weaker than that obtained after hybridization with polynucleotide probes. The probes did not hybridize with DNA from V. cholerae O1, V. cholerae non-O1 (NAG), or Y. enterocolitica. The strains of E. coli harboring the plasmid vectors of the polynucleotide gene probes were, likewise, negative in the hybridization assays.
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PMID:Cloned polynucleotide and synthetic oligonucleotide probes used in colony hybridization are equally efficient in the identification of enterotoxigenic Escherichia coli. 306 60

Strains of Vibrio cholerae O1, El Tor resistant to multiple antimicrobial agents, were isolated in Kenya between 1982 and 1985. Strains of serotype Ogawa were resistant to tetracycline, ampicillin, and trimethoprim/sulfamethoxazole. Resistance was mediated in all instances by a plasmid ca 100 mD of incompatibility group C. Based on analysis of restriction endonuclease digests, all Ogawa isolates had an identical resistance plasmid. This plasmid differed from plasmids in resistant V. cholerae O1 strains isolated in Tanzania, Nigeria, and Bangladesh. On Southern blot analysis of restriction endonuclease digests of chromosomal DNA using DNA probes there were no apparent differences between Kenyan V. cholerae O1 strains isolated before and after emergence of antibiotic resistance; however, a majority of El Tor strains isolated in other geographic areas had the same Southern blot pattern. Our data document the apparent endemicity of multiply antimicrobial resistant V. cholerae O1 strains in Kenya, and the persistence of a single unique resistance plasmid among isolates of serotype Ogawa.
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PMID:Epidemiology of antimicrobial resistant cholera in Kenya and East Africa. 319 96

The 25-kDa peptidoglycan-associated outer-membrane protein and most likely porin of Vibrio cholerae is a major immunogenic species. It has been purified by ion-exchange elution on hydroxyapatite followed by gel filtration on Bio-Gel P150 both in the presence of sodium dodecyl sulfate. This protein, of greater than 90% purity as judged by Western blotting, has been used to raise antibodies in rabbits. The antisera were then used to screen V. cholerae gene banks, constructed in Escherichia coli K12, and this has enabled us to isolate several colonies harbouring the cloned gene. The plasmids in these colonies have been designated pPM451, pPM455 and pPM472. These plasmids have a 5.3 X 10(3)-base BamHI fragment of V. cholerae DNA in common. Restriction endonuclease mapping of these plasmids has been performed and the protein identified both by Western blot analysis and in E. coli K12 minicells. The protein is not efficiently expressed in E. coli K12. It is proposed to use the name ompV to describe the structural gene, present in the cloned DNA, for this V. cholerae outer membrane protein.
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PMID:Purification of the 25-kDa Vibrio cholerae major outer-membrane protein and the molecular cloning of its gene: ompV. 398 95

The enterotoxin regions of the heat-labile and heat-stable enterotoxin (LT+ ST+) plasmid, pJY11, originating in a clinically isolated Escherichia coli strain, have been isolated as various-sized deoxyribonucleic acid (DNA) fragments by using cloning vehicles. The structure of the LT+ region and its neighboring DNA regions was studied by utilizing these recombinant plasmids. The LT+ region consisted of at least two genes, toxA and toxB, which could complement each other in trans. The toxA- and toxB-encoded polypeptides (LT subunits A and B, respectively) were identified by their immunological cross-reactivity with Vibrio cholerae enterotoxin subunit A or B. These tox genes and the promoter(s) were localized with respect to the restriction endonuclease cleavage map. The LT+ region was flanked by repeated DNA sequences (designated as beta). Another tox gen(s), encoding ST (designated as toxS), which was also flanked by inverted, repeated DNA sequences (designated as alpha), was located between one of the beta sequences and the LT+ region. These novel DNA structures (beta-alpha-toxS-alpha-toxA-toxB-beta) suggest the possibility that the LT+ region is on a transposon containing an ST transposon within the structure.
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PMID:Escherichia coli heat-labile enterotoxin genes are flanked by repeated deoxyribonucleic acid sequences. 625 52


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