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)

The occurrence of trimethoprim (Tp) resistance in salmonellas isolated from humans and water samples in Sicily between 1985 and 1988 has been investigated and the Tp resistance mechanisms have been further characterized on the basis of hybridization with probes for the dihydrofolate reductase (DHFR) genes types I, II, IV and V. Of 765 strains examined, high level (> 1000 mg/l) resistance to Tp was identified in 23 strains (3%). In 22 of these strains, such resistance was associated with resistance to sulphonamides. Six serovars with Tp-resistant strains were identified, Salmonella typhimurium (14 strains), S. enteridis (2), S. agona (2), S. mbandaka (2), S. virchow (2), S. indiana (1). In all strains with high level Tp resistance, resistance to this antimicrobial was plasmid-encoded, in most strains by plasmids with MWs ranging from 70-100 MDa. On the basis of restriction endonuclease analysis, four different categories of Tp resistance plasmids were identified in Tp-resistant strains of S. typhimurium. Hybridization with the DHFR I probe was observed in three strains of Tp-resistant S. typhimurium and two strains of Tp-resistant S. enteritidis; in contrast, in none of the strains tested was there any detectable hybridization with the probes for DHFR types II, IV and V. It is concluded that the DHFR type I resistance mechanism, common in Tp-resistant enterobacteria in many European countries, is relatively uncommon in Tp-resistant salmonellas isolated in Sicily. Furthermore, the DHFR V resistance mechanism, previously identified in strains of Shigella sonnei isolated in Sicily and associated with travellers from Sri Lanka, has not yet appeared in salmonellas in Sicily.
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PMID:Molecular characterization of trimethoprim resistance in salmonellas isolated in Sicily, 1985-1988. 748 71

Shigella sonnei is a major cause of diarrheal disease in developed as well as in developing countries. Epidemiologic studies of this organism have been limited by the lack of a simple and effective method for comparing strains. In this study, we have compared different molecular typing methods, i.e., plasmid profile analysis, restriction endonuclease analysis of plasmids, rRNA gene restriction analysis (ribotyping), pulsed-field gel electrophoresis (PFGE), and enterobacterial repetitive intergenic consensus (ERIC) sequence-based PCR (ERIC-PCR) for typing 20 clinical isolates of S. sonnei collected from six incidents of infection. PFGE and ERIC-PCR fingerprintings had the highest discriminatory power for discrimination of epidemiologically related isolates from epidemiologically unrelated strains of S. sonnei, and both gave seven distinct strain types among these isolates and the type strain of the species. Plasmid study and ribotyping produced only six and typing techniques demonstrated two distinct patterns, respectively, among these strains. All of these molecular an identical fingerprint for eight temporally related sporadic isolates. It is possible that these temporally related isolates belonged to a single bacterial clone and circulated obscurely through the community. Our results indicate that the ERIC-PCR technique represents a rapid and simple means for typing S. sonnei with a level of discrimination equivalent to that of PFGE but greater than those of plasmid profile analysis, restriction endonuclease analysis of plasmids, and ribotyping.
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PMID:Analysis of clonal relationships among isolates of Shigella sonnei by different molecular typing methods. 754 79

Restriction fragment length polymorphisms (RFLPs) of genomic DNAs from 49 clinical isolates of Shigella sonnei were analyzed by using a modified restriction endonuclease analysis procedure to investigate the genetic variability of this species. After cleavage with the restriction enzyme HaeIII or RsaI, DNA samples were electrophoresed in polyacrylamide gels and the RFLP patterns were visualized by silver staining. The results showed that among 20 strains associated with sporadic cases of infection in three Canadian provinces, 15 distinct RFLP patterns were revealed by HaeIII digestion and 12 distinct patterns were revealed by RsaI digestion. In contrast, the RFLP patterns of individual isolates within six groups of epidemiologically related isolates were identical to each other but distinct from those of unrelated isolates, and these patterns could be used to determine the genetic relationships between isolates associated with separate outbreaks of shigellosis. Our results indicate that the modified restriction endonuclease analysis technique represents a rapid, reproducible, and highly discriminatory method for the molecular typing of this species.
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PMID:Genetic variability and molecular typing of Shigella sonnei strains isolated in Canada. 791 22

A 2648-bp fragment from the P4 plasmid of Shigella sonnei strain 47 coding for the SsoII restriction endonuclease (ENase) and methyltransferase (MTase) (recognition sequence 5'-CCNGG) was sequenced. Two divergently arranged open reading frames of 905 bp for the SsoII ENase (R.SsoII) and 1137 bp for the MTase (M.SsoII) were identified. The coding regions are separated by 110 bp. The calculated M(r) of R.SsoII (35937) and M.SsoII (42887) are in good agreement with values previously obtained by in vitro transcription-translation experiments, i.e., 35 and 43 kDa for the ENase and MTase, respectively. The M.SsoII amino acid (aa) sequence revealed a considerable similarity to m5C-MTases recognizing the related sequences--M.EcoRII, M.dcm, M.MspI, M.BsuFI, M.HpaII, and M.HhaI. Surprisingly, the greatest degree of homology has been observed between the aa sequences of M.SsoII and M.NlaX, with an unidentified recognition sequence. The multiple alignment of aa sequences helps to identify the blocks of conserved aa in variable regions of MTases. These conserved aa can play a key role in target recognition. Some aspects of evolution of m5C-MTases are discussed.
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PMID:Analysis of the nucleotide and derived amino acid sequences of the SsoII restriction endonuclease and methyltransferase. 791 6

Plasmids of 70MDa from 6 Shigella sonnei isolates originating in Kaohsiung, Taiwan were found. All of them encoded resistance to chloramphenicol and tetracycline. These plasmids belonged to the H II incompatibility group. The fertility inhibition property of plasmids was the fi- type. Restriction endonuclease analysis revealed the six plasmids all had identical restriction patterns with BamH I, Hind III and Nru I. All six plasmids carried the type I chloramphenicol acetyltransferase gene and the class B tetracycline resistance gene as determined by Southern hybridization with DNA probes of various antimicrobial resistance genes. The results demonstrated that S. sonnei isolates harboured the same R plasmid.
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PMID:Characterization of R plasmids from Shigella sonnei in Taiwan. 804 80

We describe here a new approach for analyzing nucleic acid sequences using a structure-specific endonuclease, Cleavase I. We have applied this technique to the detection and localization of mutations associated with isoniazid resistance in Mycobacterium tuberculosis and for differentiating bacterial genera, species and strains. The technique described here is based on the observation that single strands of DNAs can assume defined conformations, which can be detected and cleaved by structure-specific endonucleases such as Cleavase I. The patterns of fragments produced are characteristic of the sequences responsible for the structure, so that each DNA has its own structural fingerprint. Amplicons, containing either a single 5'-fluorescein or 5'-tetramethyl rhodamine label were generated from a 620-bp segment of the katG gene of isoniazid-resistant and -sensitive M. tuberculosis, the 5' 350 bp of the 16S rRNA genes of Escherichia coli O157:H7, Salmonella typhimurium, Salmonella enteritidis, Salmonella arizonae, Shigella sonnei, Shigella dysenteriae, Campylobacter jejuni, staphylococcus, hominis, Staphylococcus warneri, and Staphylococcus aureus and an approximately 550-bp DNA segment comprising the intergenic region between the 16S and 23S rRNA genes of Salmonella typhimurium, Salmonella enteritidis, Salmonella arizonae, Shigella sonnei, and Shigella dysenteriae serotypes 1, 2, and 8. Changes in the structural fingerprints of DNA fragments derived from the katG genes of isoniazid-resistant M. tuberculosis isolates were clearly identified and could be mapped to the site of the actual mutation relative to the labeled end. Bland patterns which clearly differentiated bacteria to the level of genus and, in some cases, species were generated from the 16S genes. Cleavase I analysis of the intergenic regions of Salmonella and Shigella species differentiated genus, species, and serotypes. Structural fingerprinting by digestion with Cleavase I is a rapid, simple, and sensitive method for analyzing nucleic acid sequences and may find wide utility in microbial analysis.
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PMID:Differentiation of bacterial 16S rRNA genes and intergenic regions and Mycobacterium tuberculosis katG genes by structure-specific endonuclease cleavage. 894 Apr 59

The regulation of the Sso II restriction-modification system from Shigella sonnei was studied in vivo and in vitro . In lacZ fusion experiments, Sso II methyltransferase (M. Sso II) was found to repress its own synthesis but stimulate expression of the cognate restriction endonuclease (ENase). The N-terminal 72 amino acids of M. Sso II, predicted to form a helix-turn-helix (HTH) motif, was found to be responsible for the specific DNA-binding and regulatory function of M. Sso II. Similar HTH motifs are predicted in the N-terminus of a number of 5-methylcytosine methyltransferases, particularly M. Eco RII, M.dcm and M. Msp I, of which the ability to regulate autogenously has been proposed. In vitro, the binding of M. Sso II to its target DNA was investigated using a mobility shift assay. M. Sso II forms a specific and stable complex with a 140 bp DNA fragment containing the promoter region of Sso II R-M system. The dissociation constant (Kd) was determined to be 1.5x10(-8) M. DNaseI footprinting experiments demonstrated that M. Sso II protects a 48-52 bp region immediately upstream of the M. Sso II coding sequence which includes the predicted -10 promoter sequence of M. Sso II and the -10 and -35 sequences of R. Sso II.
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PMID:Specific binding of sso II DNA methyltransferase to its promoter region provides the regulation of sso II restriction-modification gene expression. 915 10

The nucleotide sequence of the chromosomally encoded type II ScrFI restriction/modification system from Lactococcus lactis subsp. cremoris UC503 was completed. The ScrFI restriction endonuclease (ENase) has previously been shown to specifically recognize 5' CCNGG 3' sites, cleaving after the second cytosine and the degenerate central base. The ENase gene (scrFIR; 362 bp) was located between, and co-directionally transcribed with, two formerly characterized 5-methylcytosine methyltransferase genes, which encodes proteins that independently confer protection against ScrFI digestion. scrFIR codes for a protein of 272 amino acids with a predicted molecular mass of 31470 Da, which agrees favourably with a previously estimated molecular mass of 34 kDa for this enzymes. The deduced sequence of this protein did not show any significant homology with known protein sequences, including the isoschizomeric Ssoll ENase from Shigella sonnei. The ENase gene was cloned and expressed in Escherichia coli and Lactococcus; however, no in vivo restriction of phage was observed, suggesting that expression of the ENase gene may be repressed, or that the appropriate expression signals may be absent in the cloned constructs. The ability of ScrFI to cleave non-canonically modified 5' CCNGG 3' sequences suggested that some ScrFI sites may require complex modifications to fully impair digestion by this enzyme.
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PMID:Molecular characterization of the restriction endonuclease gene (scrFIR) associated with the ScrFI restriction/modification system from Lactococcus lactis subsp. cremoris UC503. 924 16

Fifty-eight isolates of Shigella sonnei from three outbreaks in school children and eight control isolates from epidemiologically unrelated sporadic clinical infections in Taiwan were compared by antibiotic susceptibility testing and molecular typing. Antibiotic susceptibility testing showed that all strains except one sporadic isolate were multi-resistant. Ribotyping after restriction endonuclease digestion with SalI, PvuII and HindII generated the same ribosomal pattern in 65 of the 66 isolates. Plasmid profile analysis and pulsed-field gel electrophoresis (PFGE) produced eight and nine distinct patterns, respectively, and were in agreement with the epidemiological relationship of the outbreak strains. Nevertheless, some of the sporadic isolates could be discriminated only by a combination of these two methods. This study showed that plasmid profiling in combination with PFGE may be superior to ribotyping in molecular epidemiological investigations of S. sonnei.
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PMID:Molecular analysis of Shigella sonnei isolated from three well-documented outbreaks in school children. 1075 30

A strategy for the detection, identification, and differentiation of enteroinvasive Escherichia coli (EIEC) and Shigella spp. has been developed. The strategy includes (i) a multiplex PCR for the amplification of two virulence genes, i.e., iuc (222 bp) and ipaH (629 bp); (ii) amplification of the ial gene (a 1,038-bp amplicon) located within a large plasmid; and (iii) restriction fragment length polymorphism (RFLP) of the ial gene amplicon. The multiplex PCR provided three patterns. Pattern 1 (iuc-/ ipaH+) was found in 10 (67%) of 15 EIEC strains tested, pattern 2 (iuc+/ipaH-) in only 2 (4.4%) of 46 non-EIEC isolates, whereas pattern 3 (iuc+/ipaH+) was observed in all Shigella spp. and also in 5 (33%) of 15 EIEC strains tested. The pattern 3 EIEC strains were all positive for the ial gene. The PCR-RFLP of the ial gene amplicon using the endonuclease AclI was used to differentiate Shigella spp. from the EIEC strains that belonged to pattern 3. The ial gene was present in 21 (38%) of 56 and 6 (40%) of 15 Shigella spp. and EIEC strains tested, respectively. The PCR-RFLP of the ial gene amplicon divided the strains in two types. Type 1 did not contain the restriction enzyme site and was found in 6 (100%) of 6 EIEC strains, 4 (80%) of 5 Shigella boydii, and 4 (100%) of 4 Shigella dysenteriae strains tested. Type 2, which gave two fragments of 286 and 752 bp, was observed in 5 (83%) of 6 Shigella fiexneri strains and 6 (100%) of 6 Shigella sonnei strains. Detection, identification, and differentiation of Shigella spp. and EIEC were achieved by analyses of the PCR patterns and RFLP types. To our knowledge, this is the first study to demonstrate a simple and rapid method for detecting, identifying, and differentiating, at the molecular level, Shigella spp. and EIEC strains. This method will have tremendous utility as an epidemiological tool and in helping to develop policies, risk assessments, and national and international methods for Shigella spp.
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PMID:Molecular strategies for the detection, identification, and differentiation between enteroinvasive Escherichia coli and Shigella spp. 1572 63


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