Replication of the plasmid pBR322 under the control of a cloned replication origin from the single-stranded DNA phage M13.

• DOI: 10.1073/pnas.77.8.4638
• Publication Date: 2006-05-31T08:26:11Z
• Authors: J M Cleary; D S Ray
• Keywords: Replicon; In vitro recombination; T-DNA Binary system

The replication origins of viral and complementary strands bacteriophage M13 DNA are contained within a 507-nucleotide intergenic region the genome. Chimeric plasmids have been constructed by inserting restriction endonuclease fragments into plasmid pBR322. Replication these hybrid plasmids, under conditions not permissive for replicon, depends on specific segments origin presence helper virus. Thus M13-infected polA- Escherichia coli can be transformed to ampicillin resistance that functional origin. Cells drug bearing sequences contain duplex chimeric at high copy number but do accumulate significant amounts single-stranded DNA. Rare transducing phages carrying produced detected their ability transduce cells resistance. Plasmids containing 270-nucleotide fragment from gene II-proximal half produce transformants frequency nonpermissive conditions. A central Hae III fragment, III-G, nucleotide sequence coding RNA primer strand nicking site II protein, is sufficient in transformation. Additional information side III-G necessary efficient transformation