Amplification of two endo R · Hin d III-restricted fragments of the DNA of herpes simplex virus type 1
Identifieur interne : 005102 ( Main/Merge ); précédent : 005101; suivant : 005103Amplification of two endo R · Hin d III-restricted fragments of the DNA of herpes simplex virus type 1
Auteurs : N. Biswal [États-Unis] ; S. Sharma [États-Unis] ; N. C. Khan [États-Unis] ; G. A. Cabral [États-Unis] ; M. Patterson [États-Unis]Source :
- Virology [ 0042-6822 ] ; 1978.
English descriptors
- Teeft :
- Agarose, Biol, Biswal, Buoyant density, Capillary tubes, Cells transfected, Centrifugation, Circularized, Circularized fragment, Cleavage, Cleavage patterns, Cohesive ends, Cscl, Cscl density gradient centrifugation, Defective virions, Deletion mutants, Distinct sizes, Electron micrograph, Electron microscopy, Endo, Endonuclease, Equilibrium centrifugation, Ethidium bromide, Fragment, Fragment cells, Fragment ntransfected cells, Further experiments, Genome, Growth medium, Hayward, Helper, Herpes, Herpes simplex virus, Herpes simplex virus type, Herpesvirus, Higher proportions, Hind, Individual fragments, Internal inversion, Intracellular, Joint region, Ligase, Molecular weight, Molecule, Monomeric, Monomeric circles, Mutant, Nacl, Neutral sucrose density gradients, Normal size, Optical absorbancy, Other fragments, Polynucleotide, Polynucleotide ligase, Rabbit kidney cells, Reaction mixture, Reassociation, Replication, Restriction endonuclease, Right panel, Sedimentation, Simian virus, Simplex, Such fragments, Sucrose, Sucrose density gradient, Terminal fragments, Terminal sequences, Total volume, Transfected, Transfected cells, Transfection, Unique region, Viral, Viral genome, Virion, Virology, Virus type, Wilkie.
Abstract
Abstract: The 56 S DNA molecules of herpes simplex virus type 1 (HSV-1) were cleaved by restriction endonuclease Hind III (endo R · Hind III), and two of the restricted fragments were amplified in primary rabbit kidney cells by the following procedure. The restricted linear fragments were isolated after electrophoresis in 0.3% agarose gels and then treated with T4-polynucleotide ligase under conditions which favor monomeric, in contrast to concatemeric, circularization of fragments with cohesive ends. When visualized by electron microscopy, low molecular weight (<107) restricted fragments were found to be circularized with an efficiency greater than 50%, while larger fragments circularized with an efficiency of 15 to 40%. Two of the ligase-treated fragments were used to transfect rabbit kidney cells in the presence of helper infectious viral DNA. These were fragment C, molecular weight 23 × 106, belonging to the joint region of the long and short components, and fragment N, molecular weight 3 × 106, belonging to the S component of the HSV-1 DNA. Analysis of the endo R · Hind III cleavage patterns of the intracellular viral DNA coupled with reassociation kinetics indicated that these two DNA fragments replicated as monomers, dimers, and even in oligomeric forms. The maximum sedimentation coefficient of the amplified oligomeric forms, however, was lower than that of mature 56 S viral DNA.
Url:
DOI: 10.1016/0042-6822(78)90462-2
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Biswal</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030</wicri:regionArea><wicri:noRegion>Texas 77030</wicri:noRegion></affiliation></author><author><name sortKey="Sharma, S" sort="Sharma, S" uniqKey="Sharma S" first="S." last="Sharma">S. Sharma</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030</wicri:regionArea><wicri:noRegion>Texas 77030</wicri:noRegion></affiliation></author><author><name sortKey="Khan, N C" sort="Khan, N C" uniqKey="Khan N" first="N. C." last="Khan">N. C. Khan</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030</wicri:regionArea><wicri:noRegion>Texas 77030</wicri:noRegion></affiliation></author><author><name sortKey="Cabral, G A" sort="Cabral, G A" uniqKey="Cabral G" first="G. A." last="Cabral">G. A. Cabral</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030</wicri:regionArea><wicri:noRegion>Texas 77030</wicri:noRegion></affiliation></author><author><name sortKey="Patterson, M" sort="Patterson, M" uniqKey="Patterson M" first="M." last="Patterson">M. Patterson</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Virology and Epidemiology, Baylor College of Medicine, Houston, Texas 77030</wicri:regionArea><wicri:noRegion>Texas 77030</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Virology</title><title level="j" type="abbrev">YVIRO</title><idno type="ISSN">0042-6822</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1978">1978</date><biblScope unit="volume">85</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="568">568</biblScope><biblScope unit="page" to="586">586</biblScope></imprint><idno type="ISSN">0042-6822</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0042-6822</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Agarose</term><term>Biol</term><term>Biswal</term><term>Buoyant density</term><term>Capillary tubes</term><term>Cells transfected</term><term>Centrifugation</term><term>Circularized</term><term>Circularized fragment</term><term>Cleavage</term><term>Cleavage patterns</term><term>Cohesive ends</term><term>Cscl</term><term>Cscl density gradient centrifugation</term><term>Defective virions</term><term>Deletion mutants</term><term>Distinct sizes</term><term>Electron micrograph</term><term>Electron microscopy</term><term>Endo</term><term>Endonuclease</term><term>Equilibrium centrifugation</term><term>Ethidium bromide</term><term>Fragment</term><term>Fragment cells</term><term>Fragment ntransfected cells</term><term>Further experiments</term><term>Genome</term><term>Growth medium</term><term>Hayward</term><term>Helper</term><term>Herpes</term><term>Herpes simplex virus</term><term>Herpes simplex virus type</term><term>Herpesvirus</term><term>Higher proportions</term><term>Hind</term><term>Individual fragments</term><term>Internal inversion</term><term>Intracellular</term><term>Joint region</term><term>Ligase</term><term>Molecular weight</term><term>Molecule</term><term>Monomeric</term><term>Monomeric circles</term><term>Mutant</term><term>Nacl</term><term>Neutral sucrose density gradients</term><term>Normal size</term><term>Optical absorbancy</term><term>Other fragments</term><term>Polynucleotide</term><term>Polynucleotide ligase</term><term>Rabbit kidney cells</term><term>Reaction mixture</term><term>Reassociation</term><term>Replication</term><term>Restriction endonuclease</term><term>Right panel</term><term>Sedimentation</term><term>Simian virus</term><term>Simplex</term><term>Such fragments</term><term>Sucrose</term><term>Sucrose density gradient</term><term>Terminal fragments</term><term>Terminal sequences</term><term>Total volume</term><term>Transfected</term><term>Transfected cells</term><term>Transfection</term><term>Unique region</term><term>Viral</term><term>Viral genome</term><term>Virion</term><term>Virology</term><term>Virus type</term><term>Wilkie</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The 56 S DNA molecules of herpes simplex virus type 1 (HSV-1) were cleaved by restriction endonuclease Hind III (endo R · Hind III), and two of the restricted fragments were amplified in primary rabbit kidney cells by the following procedure. The restricted linear fragments were isolated after electrophoresis in 0.3% agarose gels and then treated with T4-polynucleotide ligase under conditions which favor monomeric, in contrast to concatemeric, circularization of fragments with cohesive ends. When visualized by electron microscopy, low molecular weight (<107) restricted fragments were found to be circularized with an efficiency greater than 50%, while larger fragments circularized with an efficiency of 15 to 40%. Two of the ligase-treated fragments were used to transfect rabbit kidney cells in the presence of helper infectious viral DNA. These were fragment C, molecular weight 23 × 106, belonging to the joint region of the long and short components, and fragment N, molecular weight 3 × 106, belonging to the S component of the HSV-1 DNA. Analysis of the endo R · Hind III cleavage patterns of the intracellular viral DNA coupled with reassociation kinetics indicated that these two DNA fragments replicated as monomers, dimers, and even in oligomeric forms. The maximum sedimentation coefficient of the amplified oligomeric forms, however, was lower than that of mature 56 S viral DNA.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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