Cloning and expression of the Escherichia coli replication origin in a single-stranded DNA phage.
The Escherichia coli DNA replication origin (oriC) and the adjacent asparagine synthetase gene (asnA) have been inserted into the duplex replicative form DNA of the single-stranded phage vector M13Goril. By in vitro recombination, the entire oriC asnA-containing plasmid pJS5 was inserted into M13Gori1 in both possible orientations. Both phage types transduce the asnA gene and confer upon the M13 vector the ability to replicate as a plasmid in the E. coli mutant rep3. In rep+ hosts, these phages undergo single-stranded DNA synthesis and viral morphogenesis.
D003001:Cloning, Molecular; D003090:Coliphages / Q000235:genetics; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics*; D004274:DNA, Recombinant; D004277:DNA, Single-Stranded / Q000096:biosynthesis; D004279:DNA, Viral / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics*; D009154:Mutation; D010957:Plasmids
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004262:DNA Restriction Enzymes