Date of Award
1988
Degree Type
Dissertation
Degree Name
Doctor of Philosophy
Abstract
Bacteriophage Mu integrates and replicates its genome via DNA transposition. Mu transposition during integration is non-replicative (conservative) and generates simple insertions. Transposition during the lytic cycle is replicative and amplifies the Mu genome by cointegrate production. Mu therefore, must choose between these two pathways. Infecting Mu-DNA is found associated with a coinjected, 64-kDa virion protein bound noncovalently to its ends. Characterization of the protein-DNA complex is reported here.;Antiserum was prepared against the virion 64-kDa protein and used to probe an expression library of cloned Mu DNA sequences. The Mu {dollar}N{dollar} gene was mapped to the overproducing clones by physical and genetic techniques. Partial proteolysis of the N protein produced in vitro and the 64-kDa protein isolated from the protein-DNA complex showed them to be identical. The Mu {dollar}N{dollar} gene was sequenced and a region of homology to many site specific DNA binding proteins was observed.;The transposition end product produced in vitro by the N protein-Mu DNA complex isolated from Mu infected cells was examined by neutral and alkaline agarose gel electrophoresis. The protein-DNA complex was found to form an identical strand transferred end product as the mini-Mu control plasmid. The implication of these findings for Mu integration is that in vivo, the strand transferred product is probably processed by a second nick following the initial strand transfer reaction of transposition.
Recommended Citation
Gloor, Gregory Brian, "Characterization Of The Integrative Precursor Protein-dna Complex Of Bacteriophage Mu" (1988). Digitized Theses. 1696.
https://ir.lib.uwo.ca/digitizedtheses/1696