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)

Amplification of Bacillus subtilis DNA fragments was performed in Escherichia coli using plasmid RSF2124. The main principle of isolation and cloning hybrid plasmids was described using genes of riboflavin operon as a model. Bac. subtilis DNA was treated with restriction endonuclease EcoR; followed by the agarose gel electrophoretic separation of the resulting fragments. Gels were sliced, DNA was eluted from the corresponding slices and used to transform Bac. subtilis auxotrophs rib A72, rib S110 and rib D107. DNA fraction with the molecular weight 7 . 10(6) daltons restored prototrophy of these mutants. DNA of this fraction was ligated with EcoRI treated plasmid RSF2124 DNA and used for transformation of E. coli rk-mk+. Ampicillin resistant transformants which had lost the colicin production ability, were selected. The presence of riboflavin genes within the hybrid plasmids was detected by transformation of B. subtilis auxotrophs. Three hybrid plasmids (pPR1, pPR2 and pPR3), containing a fragment of Bac. subtilis DNA with the molecular weight 6.8 . 10(-6) daltons including riboflavin operon, were selected. The analysis of the transformation activity of Bac. subtilis DNA and plasmid pPR1 DNA revealed, that there was no restriction activity of Bac. subtilis cells against plasmid DNA amplified in E. coli. Heteroduplex analysis has shown that plasmids pPR1 and pPR2 differ in the orientation of Bac. subtilis DNA fragment. DNA of these plasmids restored prototrophy of the several studied E. coli riboflavin auxotrophs.
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PMID:[Amplification of the riboflavin operon genes of Bacillus subtilis in Escherichia coli cells]. 10 60

Ampicillin-resistant strains of Shigella dysenteriae type 1 isolated in epidemics in Mexico, Central America, and Bangla Desh were examined for the presence of plasmid deoxyribonucleic acid (DNA) by gel electrophoresis. All strains contained a heterogeneous population of plasmids. Transfer experiments to Escherichia coli K-12 indicated that the ampicillin resistance determinant (Ap(r)) was located on a 5.5-megadalton (Mdal) plasmid identical in all Shiga strains examined, as judged by DNA hybridization and by its molecular properties. This 5.5-Mdal plasmid contained the ampicillin transposon (TnA) sequences. There was not a high degree of homology between the Shiga Ap(r) plasmid DNA and DNA obtained from Ap(r)Salmonella typhi strains isolated from typhoid epidemics in Mexico, previous to the dysentery outbreaks. Although low, the degree of reassociation observed indicated that probably part of the TnA sequence was present in S. typhi DNA. The DNA hybridization experiments showed, in addition, that there was a high degree of homology among Ap(r) plasmids isolated from different enterobacteria, and this identity was confirmed by restriction endonuclease activity. These results together with their similarities in molecular and replicative properties indicate that the Ap(r) plasmids, as was suggested for the Sm(r) Su(r) plasmids, possibly evolved once and then epidemiologically spread in the Enterobacteriaceae.
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PMID:Characterization of an R-plasmid associated with ampicillin resistance in Shigella dysenteriae type 1 isolated from epidemics. 32 94

We described plasmid mediated transfer of resistance to beta-lactam antibiotics between Bacteroides fragilis strains. Ampicillin-resistance was transferred from B. fragilis strain GAI-10150 to a B. fragilis strain JC-101 with a frequency of 10(-6)/input donor by a filter mating technique. A common plasmid band, named pBFKW1, was found in both the donor and the transconjugants. The plasmid was purified by an ethidium bromide-CsCl ultracentrifugation. The molecular size of the plasmid pBFKW1 which seemed to encode the beta-lactam resistance and beta-lactamase production was estimated ca. 40 kb by the analysis of endonuclease digest. Substrate profile of the enzymes derived from the donor and a transconjugant were of cephalosporinase character.
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PMID:R-plasmid mediated transfer of beta-lactam resistance in Bacteroides fragilis. 225 28

We have investigated the possibility of using restriction fragment length polymorphisms (RFLPs) to distinguish different strains of Listeria monocytogenes. Cloned DNA fragments (probes) were selected from a bacteriophage lambda gene library of L. monocytogenes (L1428). DNAs from two lambda clones consisting of a total of approximately 20kb of probe DNA were labelled with biotinylated dUTP for use in these experiments. The criteria for probe selection were the ability both to hybridise with all strains of L. monocytogenes and to reveal RFLPs. Southern blots of restriction endonuclease (Nci I) digested DNAs from test strains of L. monocytogenes were hybridized to the probe. Following washing, biotinylated probe remaining bound to the filters was detected using the Blugene reagents (Gibco/BRL). Under these experimental conditions strain-specific restriction fragments were revealed. We have examined 64 strains of L. monocytogenes belonging to serogroup 1/2 which were not apparently epidemiologically associated and 19 patterns were observed. Epidemiologically related strains gave identical patterns of restriction fragments.
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PMID:Typing of Listeria monocytogenes for epidemiological studies using DNA probes. 257 79

Native Alaskans have a high incidence of disease caused by invasive Haemophilus influenzae type b and represent an isolated population for epidemiological study. We used plasmid DNA analysis and subtyping of outer membrane proteins as markers to characterize 29 ampicillin-resistant, invasive strains and seven ampicillin-resistant, noninvasive strains of this organism from distinct geographic regions. All 36 strains produced beta-lactamase; 34 strains transferred resistance by conjugation. Seven of the 36 strains harbored detectable plasmid DNA: four had a molecular mass of 40 MDa, and three had a molecular mass of 3 MDa. Furthermore, 20 transconjugants had a similar large plasmid, and four had a similar small plasmid. Ten of 12 transconjugants with either the large, small, or undetectable plasmid DNA were able to retransfer resistance. Transformation of resistance was successful with two large plasmids. DNA-DNA hybridization studies revealed that 33 of 36 strains had DNA homology. Restriction endonuclease digestion of 10 large plasmids revealed five patterns; identity was evident within a geographic region, and similarity existed between regions. Seven restricted plasmids demonstrated an identical pattern with a small beta-lactamase probe. Ampicillin resistance in these isolates from Alaska is primarily due to a common, 40-MDa conjugative plasmid that mediates beta-lactamase production, a finding which differs from that for ampicillin-resistant plasmids isolated elsewhere in the United States. Despite variable outer membrane protein profiles of the distinct strains of H. influenzae type b, the plasmids shared significant DNA homology. It appears that a common genetic element was responsible for the dissemination of this phenotype for resistance in Alaska.
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PMID:Molecular epidemiology of plasmid-mediated ampicillin resistance in Haemophilus influenzae type b isolates from Alaska. 298 65

Recombinant DNA molecules were constructed from the plasmid pIL203 and the EcoRI-fragment of N3 plasmid containing EcoRII endonuclease and methylase genes and also a gene for resistance to sulfanilamide. The pIL203 plasmid, used as a vector, consisted of the Bam HI-EcoRI-fragment of the plasmid pBR322 conferring resistance to ampicillin and the Bam HI-EcoRI-fragment of lambda phage containing promoters, a thermosensitive mutation in the cI gene and a suppressible amber mutation in the cro gene. Ampicillin-sulfanilamide-resistant clones were selected and tested for their restriction and modification phenotype. The recombinant plasmid DNA, isolated from ApRSuR-resistant clones, which restricted and modified phage lambda imm21 with EcoRII specificity, had the EcoRI-fragment with EcoRII genes in a single orientation. The recombinant plasmid pSK323 was transferred into E. coli strains with su-, su1, su2 or su3 phenotypes. The synthesis of products of EcoRII genes by these strains grown at 37 degrees C is increased by 10--50-fold.
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PMID:Overproduction of the EcoRII endonuclease and methylase by Escherichia coli strains carrying recombinant plasmids constructed in vitro. 626 80

A map of cleavage sites for restriction endonuclease EcoRI, BamHI, HindIII, and SalI on Tn2603, a transposon encoding resistance to ampicillin, streptomycin, sulfonamide, and mercury, was constructed by an analysis of restriction cleavage patterns of plasmid pMK1.::Tn2603 and its deletion derivative. By cloning the fragments generated from pMK1.::Tn2603 with these restriction endonucleases to a pACYC184 plasmid vehicle, the regions necessary for expression of resistance were located on the restriction cleavage map of Tn2603. Ampicillin, streptomycin, and sulfonamide-resistance genes were mapped in a cluster on the region between the center and the right and the mercury-resistance gene was located to the left of the map. The final functional map of Tn2603 was compared with those of Tn4 and Tn21 and the evolutional relationships between them were discussed.
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PMID:Physical and functional mapping of Tn2603, a transposon encoding ampicillin, streptomycin, sulfonamide, and mercury resistance. 626 20

During the last 6 years in Greece, there has been a significant increase in the number of ampicillin-resistant Salmonella clinical isolates reported. In this study 23 ampicillin-resistant Salmonella strains, consecutively isolated from patients with epidemiologically unrelated cases of food poisoning, were investigated. By serotyping and phage typing, 21 of these strains were identified as Salmonella enteritidis phage type 6a, 1 was identified as Salmonella typhimurium, and 1 was identified as Salmonella saintpaul. By plasmid pattern analysis, the 21 S. enteritidis strains were further differentiated into five groups. Group I consisted of 5 strains (carrying two plasmids of ca. 38 and 34 MDa), group II consisted of 10 strains (three plasmids of ca. 38, 34, and 2.5 MDa), group III consisted of 3 strains (four plasmids of ca. 38, 34, 15, and 2.5 MDa), group IV consisted of 1 strain (five plasmids of ca. 100, 38, 34, 24, and 15 MDa), and group V consisted of 2 strains (three plasmids of ca. 100, 38, and 24 MDa). Ampicillin resistance was easily transferred to Escherichia coli and was associated with the transfer of the 34-MDa plasmid, classified in the N incompatibility group for all strains of groups I to IV, and with the transfer of the 100-MDa plasmid for the group V strains. EcoRI restriction endonuclease digestions showed an extensive homology among the 34-MDa conjugative R plasmids. Hybridizations of the EcoRI restriction fragments of the 34-MDa plasmids with a TEM-type probe revealed the locus of the beta-lactamase gene to be situated on a ca. 6.6-MDa fragment, common in all plasmids. These results indicate that ampicillin resistance in Greece is due to the spread of a limited number of clones of S. enteritidis phage type 6A, carrying related 34-MDa R plasmids. Work is in progress to obtain a better understanding of ampicillin resistance in S. enteritidis.
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PMID:Molecular epidemiology of ampicillin-resistant clinical isolates of Salmonella enteritidis. 805 Dec 61

Using massive cDNA sequencing, proteomics and customized computational biology approaches, we have isolated and identified the most abundant secreted proteins from the salivary glands of the sand fly Lutzomyia longipalpis. Out of 550 randomly isolated clones from a full-length salivary gland cDNA library, we found 143 clusters or families of related proteins. Out of these 143 families, 35 were predicted to be secreted proteins. We confirmed, by Edman degradation of Lu. longipalpis salivary proteins, the presence of 17 proteins from this group. Full-length sequence for 35 cDNA messages for secretory proteins is reported, including an RGD-containing peptide, three members of the yellow-related family of proteins, maxadilan, a PpSP15-related protein, six members of a family of putative anticoagulants, an antigen 5-related protein, a D7-related protein, a cDNA belonging to the Cimex apyrase family of proteins, a protein homologous to a silk protein with amino acid repeats resembling extracellular matrix proteins, a 5'-nucleotidase, a peptidase, a palmitoyl-hydrolase, an endonuclease, nine novel peptides and four different groups of proteins with no homologies to any protein deposited in accessible databases. Sixteen of these proteins appear to be unique to sand flies. With this approach, we have tripled the number of isolated secretory proteins from this sand fly. Because of the relationship between the vertebrate host immune response to salivary proteins and protection to parasite infection, these proteins are promising markers for vector exposure and attractive targets for vaccine development to control Leishmania chagasi infection.
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PMID:Identification of the most abundant secreted proteins from the salivary glands of the sand fly Lutzomyia longipalpis, vector of Leishmania chagasi. 1537 79

Clindamycin-treated hamsters are predictably susceptible to infection with pathogenic strains of Clostridium difficile. This animal model parallels most of the important aspects of human C. difficile associated disease (CDAD). In humans, almost any antibiotic may precipitate CDAD, but clindamycin, ampicillin and second-and third-generation cephalosporins are implicated most often. We studied the effect of ampicillin and ceftriaxone compared to clindamycin on the susceptibility of hamsters to challenge with C. difficile strain designated B1 by restriction endonuclease typing, an epidemic strain from one hospital. Hamsters were highly susceptible to CDAD following a single dose of clindamycin (30 mg/kg orogastrically) from 1 to 4 days when challenged with 100 colony-forming units (CFU) of spores of epidemic CD strain B1. Ampicillin was given orogastrically at 60 mg/kg to groups of three hamsters that were challenged with 10000 CFU of CD strain B1 spores on days 1-4 following ampicillin. Hundred percent CDAD mortality occurred in all groups on each challenge day. Ceftriaxone, given intraperitoneally at 60 mg/kg, induced susceptibility to CDAD for a more limited time course and at a higher CD inoculum, producing 100% mortality when hamsters were challenged with 10000 CFU of CD strain B1 on day 1 following ceftriaxone, 33% mortality at day 2, and no CDAD when challenged on days 3 and 4 following ceftriaxone. Hamsters are susceptible to CD infection for at least 4 days following ampicillin and clindamycin, but ceftriaxone has a shorter duration of susceptibility.
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PMID:Susceptibility of hamsters to human pathogenic Clostridium difficile strain B1 following clindamycin, ampicillin or ceftriaxone administration. 1688 94


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