Replication and plasmid-bacteriophage recombination I. Marker rescue analysis》.pdf

VIROLOGY 115, 223-236 (1981) Replication and Plasmid-Bacteriophage Recombination I . Marker Rescue Analysis RICHARD D. SMITH' AND ROBERT C. MILLER, JR. 2 Departments of Microbiology and Medical Genetics, University geBritish Columbia, Vancouver, British Columbia V6T IW5, Canada February 20, 1981 ; accepted July 1, 1981 Received Recombinant DNA plasmids of pBR322 and T7 DNA were constructed and tested for the presence of various T7 genes. A plasmid containing part of T7 gene 5 (DNA poly- merase) was used to transform various strains of Escherichia coii, including a dna B mutant. [3 HTrhymidine incorporation and a new method of copy number analysis showed that plasmid DNA was not degraded after infection by T7 as is the E coil host DNA . Marker rescue between recombinant T7 plasmids and mutant infecting bacteriophage was quantitated by determining the percentage of wild-type progeny phage produced after infection . Replication of the plasmid DNA and infecting phage DNA was controlled independently by a dna B mutation in the host and by gene 5 mutations in the phage, respectively. Marker rescue frequencies decreased slightly, if either plasmid or phage replication was blocked . However, marker rescue dropped below detectable levels, if nei- ther the plasmid nor the phage could replicate . These results show clearly that replication plays a role in plasmid-phage recombination, and possible roles for replication in this process are discussed. INTRODUCTION bility of concatemers (Frohlich et al.,