Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

A typing method was developed for Neisseria meningitidis serogroup A by analysis of restriction fragment length polymorphisms (RFLP) of the class 1 outer membrane protein gene (porA). By using appropriate primers, an approximately 1,116-bp fragment of the porA gene was amplified by PCR and then was digested with the restriction endonuclease MspI. The digestion products were separated on 10% polyacrylamide gels and were stained with silver. One hundred three clinical isolates of group A N. meningitidis from 17 provinces of China collected over a 26-year period were analyzed. Results of MspI-generated RFLP profiles of PCR-amplified porA genes were compared with those obtained by conventional serosubtyping. There was a band of about 400 bp common to all strains examined, and the 103 strains of serogroup A resulted in 22 unique RFLP patterns. The differences in bands could be observed mainly in the range of 120 to 280 bp. The smaller fragments were useful in distinguishing meningococci with the same serosubtype. Three epidemic periods were characterized by the presence of three distinct genotypes (a1, a2, and a3), accounting for 74.5% of the strains examined (3.88, 26.21, and 44.66%, respectively). Three predominant RFLP patterns were correlated epidemiologically with cycles of epidemic meningococcal meningitis and were well-matched to the predominant serosubtypes (P1.9, P1.7, 10, and P1.9) that presented at the same prevalence cycles. The genotyping yielded information that allowed strains from one epidemic to be distinguished from those from another that would have been indistinguishable if only serotyping and serosubtyping were available. Therefore, the PCR-RFLP typing method was very useful in the epidemiologic investigation of group A meningococcal meningitis.
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PMID:Typing Neisseria meningitidis by analysis of restriction fragment length polymorphisms in the gene encoding the class 1 outer membrane protein: application to assessment of epidemics throughout the last 4 decades in China. 771 7

The application of a diagnostic and genotyping technique based on the polymerase chain reaction (PCR) to the study of trachoma epidemiology in the Gambian village of Jali is reported. PCR based on the major outer membrane protein (MOMP) gene of Chlamydia trachomatis appears to be more sensitive than either isolation or antigen detection by enzyme immunoassay; it had a specificity of 95% and sensitivity of 51% against clinical signs. PCR genotyping identified genotypes A and B of Chlamydia trachomatis circulating in Jali. Sequencing revealed a Pst1 restriction endonuclease site in the amplified MOMP gene of some B strains but not others; Pst1 digestion of the PCR product proved an easy method of distinguishing these strains. The distribution of serotypes and B strain variants shows a significant degree of household clustering (p < 0.001). PCR based genotyping combined with strain typing provides a new and powerful epidemiological tool for the study of transmission events in trachoma.
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PMID:Molecular epidemiology of trachoma in a Gambian village. 784 72

Chlamydia trachomatis serovar was determined by analysis of restriction fragment length polymorphism in the gene encoding the major outer membrane protein (MOMP) from 435 urogenital specimens. Of the specimens, 254 grew < 25 inclusions and 14 were negative in culture. Although previous studies defined serovar by epitopes or sequences representing only the four variable domains in MOMP, restriction endonuclease sites characteristic for each serovar not only within but also outside these variable domains were cataloged in this study. Novel serovars that grew poorly or not at all in vitro were not observed, and all samples proved similar or identical to one of the 15 known serovars. There was no significant difference in proportions of serovars between men and women. In women, F serovars were more frequently observed in infections with few inclusions in culture, whereas B group serovars predominated when many inclusions were observed.
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PMID:Chlamydia trachomatis serovars in 435 urogenital specimens typed by restriction endonuclease analysis of amplified DNA. 810 52

Plasmid analysis, restriction endonuclease analysis, antimicrobial susceptibility testing, biotyping, phage typing and outer membrane protein electrophoresis were used to study an outbreak of Salmonella typhimurium infection at a newborn nursery. Seven out of the 12 neonates had positive blood cultures for S. typhimurium, and 2 of them died of severe sepsis. Thirty epidemic strains of S. typhimurium belonging to phage type 12 had the same plasmid profiles (98.0, 6.7 and 3.8 Kb) and identical restriction digest patterns (23.0, 20.4, 15.0, 9.6, 8.2, 7.4, 5.8, 4.3, 3.8, 2.0 and 1.8 Kb) which were different from those of the 2 non-epidemic strains. Laboratory data suggested that the source of the infection was the index patient's mother who had a slight diarrhea; the mode of transmission was most likely due to the transfer of organisms from infant to infant by the contaminated hands of nurses during milk feeding.
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PMID:Molecular epidemiologic study of an outbreak of Salmonella typhimurium infection at a newborn nursery. 822 93

When the present chlamydial classification was established it was recognized that a wide variety of types were contained within the arbitrary designation Chlamydia psittaci. Early workers relied mostly on observations of growth characteristics to differentiate the types of C. psittaci isolated from a wide range of different hosts. The differences between isolates were confirmed serologically using a variety of tests of which the most sensitive was the micro-immunofluorescence (MIF) test which was able to recognize nine immunotypes among the mammalian isolates alone. This approach has recently been improved by the use of monoclonal antibodies in the MIF test which has confirmed most of the mammalian immunotypes and divided the avian strains into four groups. Studies on the nucleic acid of C. psittaci isolates show clear differences in the size distribution of DNA fragments produced by restriction endonuclease digestion of the genomes of the various types. Most importantly, studies of DNA/DNA homologies showed that at least four of the types identified by biological, serological and restriction endonuclease tests were sufficiently different to be considered separate species. Most recently, attention has been focused on DNA sequence comparisons of C. psittaci genes amplified by the polymerase chain reaction (PCR). The usual target has been the major outer membrane protein gene for which much sequence information is now available. The combination of PCR and MIF with monoclonals has provided a set of practical techniques with which all chlamydial isolates can be detected and typed with relative ease. It is likely that these developments will lead to the reclassification of the genus and, hopefully, a rapid increase of our understanding of the diseases caused by C. psittaci.
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PMID:Typing Chlamydia psittaci--a review of methods and recent findings. 829 58

A genomic library derived from a virulent isolate of Legionella pneumophila was constructed in Escherichia coli JM 83 using the cloning vector pUC19. The clones were screened by filter immunoassay using L. pneumophila rabbit polyclonal antisera and in the absence of in situ bacterial lysis one such clone, LP 116, expressed L. pneumophila-specific antigens on the surface of E. coli. Restriction endonuclease digest analysis and agarose gel electrophoresis revealed a fragment measuring approximately 750 bp. Southern hybridization confirmed that the fragment was L. pneumophila DNA. Sequencing data showed that the fragment was 810 bp in length with an open reading frame (ORF) of 678 bp. The outer-membrane profiles of the E. coli parent, the L. pneumophila DNA-contributing strain and clone LP 116 were compared by SDS-PAGE. A protein of 25 kDa was found in outer-membrane preparations of both the clone LP 116 and L. pneumophila but not in E. coli JM 83. This was in agreement with the molecular mass of the deduced peptide of the mature protein. Immunoblots using L. pneumophila-specific polyclonal antiserum confirmed that this 25 kDa outer-membrane protein (OMP) was a L. pneumophila polypeptide. Both direct immunofluorescence assay and immunoblots using the commercially produced monoclonal antibody specific for the common antigen of the major outer-membrane protein (MOMP) confirmed that the 25 kDa protein produced by LP 116 was involved with the MOMP complex. The gene encoding this protein has been designated ompM.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Cloning, nucleotide sequence and expression in Escherichia coli of a gene (ompM) encoding a 25 kDa major outer-membrane protein (MOMP) of legionella pneumophila. 840 14

Two different nucleic acid precursor utilization patterns were obtained for five avian isolates of Chlamydia psittaci. Three of the isolates behaved in a manner similar to that previously described, showing total dependency on the host cell for ribonucleoside triphosphates and being unable to utilize medium-supplied thymidine. In contrast, the other two isolates were incapable of taking pyrimidine ribonucleotides from the host cell and they could efficiently utilize medium-supplied thymidine. These unusual isolates were resistant to 5-fluorouridine while the other three isolates were sensitive. Of the five isolates only 6BC was sensitive to sulfonamides. The five isolates were divided into two groups by comparing the AluI restriction endonuclease patterns obtained following digestion of the major outer membrane protein (OMP1) gene, amplified by the polymerase chain reaction. The OMP1 genotyping results were confirmed by serotyping.
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PMID:Diversity in nucleotide acquisition by antigenically similar Chlamydia psittaci of avian origin. 851 20

Immunoblots using bovine antibody against Haemophilus somnus as the primary antibody consistently identified 31-, 40- and 78-kDa proteins in Sarkosyl-insoluble extracts of H. somnus. A genomic library of H. somnus 8025 DNA was constructed in plasmid pUC19, and 45 recombinants expressed proteins which were recognized by bovine antiserum in Western blots (immunoblots). Ten of the recombinants expressing a 31-kDa protein caused the lysis of bovine erythrocytes. Restriction endonuclease mapping indicated that the hemolytic recombinants shared an approximately 1.7-kb BglII fragment. Southern blot analysis using the BglII fragment as a probe revealed homology among the recombinants and the presence of an identically sized BglII fragment in the chromosome of all H. somnus isolates tested. Sequence analysis indicated the presence of an 822-bp open reading frame within the 1.7-kb BglII fragment. Deletion of this open reading frame resulted in the loss of hemolytic activity and protein expression in recombinant Escherichia coli, suggesting the possible role of the 31-kDa protein as a hemolysin. An amino acid sequence deduced from the DNA sequence shared homology with outer membrane protein A of E. coli, Salmonella typhimurium, and Shigella dysenteriae, with P6 of Haemophilus influenzae, and with PIII of Neisseria gonorrhoeae. An amino acid analysis of the recombinant 31-kDa protein agreed with the amino acid composition deduced from the DNA sequence.
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PMID:Cloning and sequencing of the gene encoding a 31-kilodalton antigen of Haemophilus somnus. 851 83

Analysis of the genomic DNAs of chlamydial isolates from sheep, cattle, and pigs was performed by Southern blot hybridization and by restriction endonuclease (RE) profiling of DNA amplified by PCR. The hybridization probes were derived from whole genomic DNA, the major outer membrane protein (MOMP) gene, the 16S rRNA gene, and an avian Chlamydia psittaci isolate plasmid. The PCR analysis used targets in the MOMP gene, the 16S rRNA gene, and the 60-kDa cysteine-rich protein gene. Together, the results showed that although there was considerable heterogeneity in the DNA sequence in the MOMP gene region, all the isolates had the same underlying total genomic RE profiles and yielded identical RE profiles for the rRNA and 60-kDa-protein gene regions. Most of the isolates were found to hybridize with the plasmid probe. Comparison of the MOMP sequence of one of the isolates (P787) with that of a known Chlamydia pecorum strain together with the results of the RE analyses allowed the conclusion that the isolates should all be classified within this new species.
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PMID:Analyses of the genomes of chlamydial isolates from ruminants and pigs support the adoption of the new species Chlamydia pecorum. 857 3

We have analyzed 81 isolates of Mycoplasma agalactiae from four different regions of Italy between 1990 and 1995 in order to identify antigenic differences through SDS-PAGE and Western blotting and chromosomal DNA restriction endonuclease cleavage pattern differences. Antigenic variability in M. agalactiae isolates was investigated analyzing hydrophobic membrane protein fractions by immunoblotting using pooled sheep antiserum from naturally infected sheep. Large restriction fragments obtained cleaving genomic DNAs with SmaI, NruI, SalI, XhoI, BssHII and KpnI were analyzed by pulsed field gel electrophoresis. Genetic analysis indicates that isolates are all similar without intraspecific differences. This homogeneity was confirmed by immunoblotting: 80 and 50 kDa antigens are present in all strains analyzed.
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PMID:Comparison of Mycoplasma agalactiae isolates by pulsed field gel electrophoresis, SDS-PAGE and immunoblotting. 883 80


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