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)

Streptomyces antibioticus produces a strong endo-DNase which is located between the cytoplasmic membrane and the cell wall. All DNA substrates assayed, including the chromosomal DNA of this species and several bacteriophage DNAs, were completely degraded in vitro by the enzyme. The rate of synthesis of the nuclease depended on the growth medium. In NBG medium, in which the enzyme is not produced, the size of lytic plaques of several actinophages was larger than that in GYM or GAE medium, in which synthesis of the nuclease takes place late in growth. In addition, one of the phages assayed, phi A6, showed a diminution of its efficiency of plating in GYM medium with respect to that in NBG medium; another phage, phi A9, grew in NBG medium but not in the other two media. It is postulated that the presence of the host nuclease, together with the capability of the particular phage to absorb on S. antibioticus of different growth phases, determines the efficiency of growth and the plaque size of the phages on productive media. This hypothesis was confirmed when the growth of phi A6 and phi A9 in a mutant of S. antibioticus lacking the endonuclease activity was analyzed. It is concluded that the enzyme can assume, under some circumstances, a role in in vivo restriction.
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PMID:An exocytoplasmic endonuclease with restriction function in Streptomyces antibioticus. 283 Feb 37

Several clinical varicella-zoster virus isolates obtained during testing of a live varicella vaccine had DNA restriction fragment patterns resembling neither vaccine nor wild-type virus [Gelb et al., J Infect. Dis. 155, 633-640, 1987]. One explanation for these isolates was recombination in vivo. To determine if such recombination is likely, two strains of varicella-zoster virus, distinguishable by restriction endonuclease fragment size differences (wild-type strain EF and the OKA vaccine strain), were grown together in tissue culture. After three passages, the mixed infection virus was plaque-purified. DNA from about 13% of the plaque-purified isolates had one or more BglI fragments found in neither parental virus. Hybridization studies showed that isolates containing one of the new BglI fragments were recombinants of the two parental strains. The BglI restriction fragment pattern of these recombinants resembled those of the unusual varicella isolates from individuals either vaccinated with the live attenuated OKA varicella vaccine and later exposed to natural varicella, or simultaneously exposed to both a recent recipient of the vaccine and natural varicella.
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PMID:Recombination in tissue culture between varicella-zoster virus strains. 283 29

A herpes simplex virus (HSV) intertypic recombinant (RE6) has been previously shown to be non-neurovirulent following direct intracranial inoculation of mice. An in vitro recombination/in vivo selection strategy was employed to further characterize the gene or genes responsible for the avirulence of this mutant. It was found that RE6 could be converted to a lethal virus by incorporation of wild-type HSV-1 sequences mapping between 0.698 and 0.721 map units. Restriction endonuclease and Southern blot analysis revealed that viral recombinants which incorporated at least part of the cloned 17syn+ sequences were selected by passage in mouse brains in vivo. The recombinants generated with this fragment were at least 50-fold more neuroviurlent than RE6, as determined by the plaque forming unit to lethal dose 50% assay. Further, they displayed a significant increase in ability to replicate in mouse brain tissue following intracranial inoculation. However, these recombinants did not display a replication advantage over RE6 in cultured mouse cells at 38.5 degrees C. Thus, the defect present within this region of the RE6 genome specifically affects replication in the nervous system. In the accompanying paper we analyze the RNA transcription and DNA sequence in this portion of the RE6 and parental strain genomes.
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PMID:Partial rescue of herpes simplex virus neurovirulence with a 3.2 kb cloned DNA fragment. 285 24

The analysis of 23 clinical isolates of herpes simplex virus type 1 (HSV-1) showed that 15 of 15 isolates that had undergone a few passages in tissue culture (fresh isolates) and two of eight isolates that had never been passaged (new isolates) were composed of a mixed population with respect to plaque morphology in Vero cells. Cloning and characterization of 10 large plaque viruses (L variants) and nine small plaque viruses (S variants), obtained from seven different isolates, showed the following. BamHI DNA restriction patterns of the L and the S variants from a single isolate differed only with respect to the electrophoretic mobility of the fragments that contain reiteration of specific sequences; they did not differ regarding the presence or the absence of restriction endonuclease cleavage sites. The L and S variants differed with respect to the electrophoretic profiles of infected cell glycoproteins, thermosensitivity of growth and plaquing efficiency at 39 degrees C, and, at least in the case of the two couples of variants that we tested, pathogenicity for the mouse. The hypothesis that the L variants might arise from the S variant during in vivo replication is discussed.
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PMID:Study of herpes simplex virus type 1 populations obtained from recurrences and primary infections. 298 77

The components of the cell cycle for a feline embryo cell line were defined. Thymidine (6mM)-supplemented medium reversibly arrested cells 1 h into the S phase of the cell cycle and was used in a double blocking procedure to synchronize cells to the early S phase. The kinetics of feline panleukopenia virus replication in synchronized cells was studied by using (i) inclusion body formation, (ii) a plaque assay for cell-associated and cell-free virus under one-step growth conditions, (iii) an enzyme immunoassay for viral protein, (iv) electron microscopy of infected cells, and (v) the detection and identification of viral replicative form DNA by restriction endonuclease analysis. Parallel studies by each of these procedures of the replication of feline panleukopenia virus in cells in which a 6 mM thymidine block was maintained indicated that parvovirus replicated with essentially similar kinetics in both unblocked, synchronized cells and in cells in which the block was maintained. Accordingly, a 6 mM thymidine-supplemented medium, although it effectively blocks cellular DNA synthesis, does not block the replication of parvovirus.
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PMID:Feline panleukopenia virus replicates in cells in which cellular DNA synthesis is blocked. 298 22

Equine herpesvirus type 3 (EHV-3) DNA, isolated from purified virions of the large-plaque strain, was digested with the restriction endonucleases XbaI, Bg/II, EcoRI, and HindIII. Several lines of evidence indicated that the DNA extracted from purified virions was composed of long (L) and short (S) components and was present as two isomeric forms, P and IS. The evidence included: (i) after electrophoresis on agarose gels, the summed molecular weights of the digestion products exceeded that expected from intact, unit size DNA; (ii) quantitative measurements of radioactivity (molar ratios) indicated 'minor bands' (0.5 M) interspersed among the major (1.0 M) bands; and (iii) a brief digestion with lambda-5'-exonuclease, prior to digestion with restriction endonuclease, resulted in the loss of some submolar and molar ratio bands, indicative of three termini. A preliminary fragment linkage map of the XbaI digestion products revealed EHV-3 DNA to contain only one recognition site in the unique sequence of the S component. From this linkage map, the size of the S component was deduced to be (22.3 +/- 5) X 10(6) molecular weight.
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PMID:Molecular pathogenesis of equine coital exanthema: restriction endonuclease digestions of EHV-3 DNA and indications of a unique XbaI cleavage site. 298 21

The serological and biological properties of the type 2 plaque-producing agent (PPA) derived from the Cal-1 strain of Marek's disease virus (MDV) were compared with those of reference strains of the three serotypes of MDV and herpesvirus of turkeys (HVT) groups; namely JM, HPRS-24 strains of MDV and the FC-126 strain of HVT for serotype 1, 2, and 3, respectively. By agar gel precipitation (AGP), indirect fluorescent antibody and virus neutralization tests, type 2 PPA was related but not identical to the FC-126 strain. By the AGP test, type 2 PPA showed a poor ability to synthesize B antigen and the A antigen was different from that of strain FC-126. To compare the virological characteristics of type 2 PPA with the reference strains, the release of cell-free virus into supernatants of infected cell cultures and titers of cell-free virus extracted sonically from infected cell cultures were examined. Cell-free type 2 PPA virus was easily detected in the supernatants and extracted from infected cell cultures. These properties were similar to reference strains of serotype 2 and 3. Next, the structural similarities of viral DNAs of type 2 PPA and strain FC-126 were examined by Southern blot hybridization. The restriction endonuclease-cleavage patterns of DNA of type 2 PPA were very similar but not identical to those of the FC-126 strain. In chickens inoculated with type 2 PPA, splenic lymphocytes supported a non-productive latent infection as did also those from chickens inoculated with the FC-126 or HPRS-24 strains. From these results, type 2 PPA appears to belong to serotype 3 of MDV and HVT groups. The origin of type 2 PPA is discussed.
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PMID:A comparison of the biological properties of type 2 plaque-producing agent derived from the Cal-1 strain of Marek's disease virus with other related viruses. 301 31

Mycophenolic acid, an inhibitor of purine metabolism, was shown to block the replication of vaccinia virus in normal cell lines. This observation led to the development of a dominant one-step plaque selection system, based on expression of the Escherichia coli gpt gene, for the isolation of recombinant vaccinia viruses. Synthesis of xanthine-guanine phosphoribosyltransferase enabled only the recombinant viruses to form large plaques in a selective medium containing mycophenolic acid, xanthine, and hypoxanthine. To utilize the selection system efficiently, we constructed a series of plasmids that contain the E. coli gpt gene and allow insertion of foreign genes into multiple unique restriction endonuclease sites in all three reading frames between the translation initiation codon of a strong late promoter and synthetic translation termination sequences. The selection-expression cassette is flanked by vaccinia virus DNA that directs homologous recombination into the virus genome. The new vectors allow high-level expression of complete or partial open reading frames and rapid construction of recombinant viruses by facilitating the cloning steps and by simplifying their isolation. The system was tested by cloning the E. coli beta-galactosidase gene; in 24 h, this enzyme accounted for approximately 3.5% of the total infected-cell protein.
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PMID:Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors. 313 Apr 92

The phage insensitivity of Streptococcus lactis KR5 was evaluated for its possible linkage to plasmid DNA. This strain possessed plasmids of 40, 29, 26, 21, 16.5, 10.5, 7.8, and 1.5 Mdal. Plasmid curing using novobiocin resulted in derivatives with increased sensitivity to prolate-headed phage, suggesting the involvement of plasmid DNA in phage insensitivity. Transformation of S. lactis LM0230 protoplasts with the KR5 plasmid DNA pool produced transformants containing a plasmid of about 27 Mdal. These erythromycin-resistant transformants were lactose-positive phage-sensitive or were lactose-negative and exhibited a reduced sensitivity to phage. Agarose gel electrophoresis and restriction endonuclease digestion analysis showed the 27-Mdal plasmid band to be composed of two distinct plasmids of 26 Mdal (pBF61) and 29 Mdal (pBF62), which coded for reduced phage sensitivity and lactose-positive phenotypes, respectively. The mechanisms of reduced phage sensitivity encoded by pBF61 included a restriction/modification system and a mechanism that resulted in reduced plaque size independent of incubation temperature. These results further support the involvement of plasmid DNA in the mechanisms for reduced phage sensitivity in dairy streptococci.
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PMID:Plasmid-mediated reduced phage sensitivity in Streptococcus lactis KR5. 313 85

A human cDNA library was constructed utilizing RNA isolated from cultured skin fibroblasts. Recombinant clones containing elastin sequences were identified by plaque hybridizations with previously characterized human placental elastin cDNAs. Seven positive recombinant clones with inserts of approximately 3.2-2.2 kb were isolated. Characterization of the clones by restriction endonuclease analysis and dot-blot hybridizations with exon-specific synthetic oligonucleotides demonstrated considerable variability in the primary nucleotide sequence. Dideoxy nucleotide sequencing confirmed this finding. The variability is most likely a result of alternative splicing of exons from the primary elastin transcripts. The two largest clones contained approximately 1 kb of 3' untranslated sequence and approximately 2.2 kb of translated sequence encoding 730 amino acids. Six amino acids, encoded by exon 12A, have not been previously noted in human elastin cDNAs. In addition, these human skin fibroblast clones contained a 49 bp 5' untranslated sequence. These results demonstrate that there is considerable variability in the processed nucleotide sequence of the elastin mRNAs. These transcripts may code for isoforms of tropoelastin with different biologic properties.
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PMID:Cloning of full-length elastin cDNAs from a human skin fibroblast recombinant cDNA library: further elucidation of alternative splicing utilizing exon-specific oligonucleotides. 317 Dec 21


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