Replication of Duplex DNA of Phage φ X174 Reconstituted with Purified Enzymes
Abstract
Replication of the covalently closed duplex replicative form (RF) of phage phi X174 DNA has been achieved by coupling two known enzyme systems: (i) synthesis of viral strand circles (SS) from RF, and (ii) conversion of SS to nearly complete RF (RF II). In this coupled system, activated RF (gene A . RF II complex) was a more efficient template and generated as many as 10 RF II molecules per RF input, at a rate commensurate with SS synthesis. The 11 proteins required for the two component systems were all needed in the coupled RF duplication system; no new factors were required. Single-stranded DNA binding protein was needed for RF duplication at only 4% the level needed in its stoichiometric participation in SS synthesis. In addition to RF II, more complex replicative forms appeared late in the reaction, and their possible origin is discussed.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- June 1980
- DOI:
- 10.1073/pnas.77.6.3322
- Bibcode:
- 1980PNAS...77.3322A