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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Zinder, N. D. Dotto, G. P. |
| Abstract | One of the most common sites used for cloning in the filamentous phages f1, fd, and M13 lies within the phage 'functional origin,' a sequence of 140 nucleotides that is required for phage replication. Even small insertions (four nucleotides) at this location severely reduce origin function. Secondary trans-acting mutations in the phage genome are necessary to restore efficient replication. One of these mutations, present in one of our cloning vectors, R218, has been fully characterized. It consists of a regulatory mutation within gene V that leads to a marked increase in the intracellular level of the phage gene II protein, the 'initiator' of viral replication. Increased gene II protein production is sufficient to reduce the minimal sequence required for a functional origin to only 40 nucleotides, while the remaining 100 (containing the cloning site) become entirely dispensable. The general implications of these findings are discussed. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 5 |
| Volume Number | 81 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1984-05-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Cloning, Molecular Coliphages Genetics DNA Replication Escherichia Coli Peptide Initiation Factors DNA Restriction Enzymes Plasmids Virus Replication 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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