A simple method for purifying the gene A plus A* proteins of bacteriophage (PHI)X174 is presented. The purified preparation was able to stimulate (PHI)X DNA replication in vitro and was able to nick supercoiled (PHI)X RFI to produce unit-length linear and circular strands of DNA. The kinetics of the nicking reaction were more consistent with a stoichiometric than with a catalytic mechanism. Under non-denaturing conditions the presumably globular A and A* proteins sedimented with apparent molecular weights of 110,000 and 90,000 daltons. A* was unable to nick supercoiled (PHI)X RFI DNA in the absence of A, but participated in the nicking reaction when A was present and both A and A* were found covalently bound to the 5'-end of the resulting nicked DNA strands. The A protein is truly cis-acting in vivo. Relaxation complexes like those observed with the E. coli colicinogenic factors were not found during (PHI)X semi-conservative DNA replication.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.68608 |
| Date | January 1981 |
| Creators | Dubeau, Louis. |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Biochemistry) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 000137644, proquestno: AAINK54781, Theses scanned by UMI/ProQuest. |
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