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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Edenberg, H. J. Waqar, M. A. Huberman, J. A. |
| Abstract | We have developed two subnuclear systems for synthesis of DNA of simian virus 40 in vitro. We prepare chromatin from infected cells by the method of Hancock [(1974) J. Mol. Biol. 86, 649-663]; these 'chromatin bodies' can be disrupted and large debris can be pelleted, leaving a supernatant ('soluble system'). Both chromatin bodies and the soluble system incorporate deoxyribonucleoside triphosphates into nucleoprotein complexes that contain simian virus 40 DNA. The DNA labeled in short pulses sediments in neutral sucrose gradients slightly faster than mature simian virus 40 DNA, as expected for replicating intermediate. When rebanded in alkaline sucrose gradients, about half of the radioactivity is found in short strands (200-300 nucleotides) and half in longer strands (up to full viral size). When these systems are supplemented with a cytoplasmic preparation from HeLa cells, synthesis is stimulated about 5-fold, and the short strands are converted into strands of up to full viral length as well as into covalently closed circles. These subnuclear DNA-replicating systems should be useful for biochemical fractionation and characterization of some of the proteins required for DNA replication. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 12 |
| Volume Number | 73 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1976-01-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Chromatin Metabolism DNA Replication DNA, Viral Biosynthesis Simian Virus 40 Virus Replication Cell Nucleus DNA, Circular Kinetics Models, Biological Molecular Weight Solubility Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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