RRC ID 29740
著者 Manis JJ, Kline BC.
タイトル Restriction endonuclease mapping and mutagenesis of the F sex factor replication region.
ジャーナル Mol Gen Genet
Abstract The plasmids pSC138 and pML31 each contain the EcoRI-generated f5 replicator fragment of the conjugative plasmid F in addition to an EcoRI fragment encoding antibiotic resistance: ampicillin resistance derived from Staphylococcus aureus in pSC138 and kanamycin resistance from Escherichia coli in pML31. We have mapped one HindIII and two BamHI restriction sites in the f5 region of these plasmids and one HindIII site in the antibiotic resistance region of each plasmid. The HindIII site in the Km region of pML31 occurs in the kan gene whereas the HindIII site in the Ap region of pSC138 appears to occur in an area important for the regulation of beta-lactamase production. By means of in vitro recombinant DNA manipulation of plasmids pML31 and pSC138, we have shown that approximately 1.9 X 10(6) daltons of the 6.0 X 10(6) dalton f5 fragment can be deleted without disrupting plasmid stability. In addition, we have used these same techniques to isolate a novel F-controlled Ap plasmid cloning vehicle which contains a single restriction site for each of the enzymes EcoRI, HindIII, and BamHI. This cloning vehicle has been linked via either its EcoRI or HindIII site to a ColE1 plasmid replicon to yield stable recombinants.
巻・号 152(3)
ページ 175-82
公開日 1977-4-29
DOI 10.1007/BF00268815
PMID 327274
MeSH Conjugation, Genetic* DNA Replication* DNA Restriction Enzymes DNA, Recombinant Endonucleases Escherichia coli F Factor* Genes* Genetic Linkage R Factors
リソース情報
原核生物(大腸菌) ME6228 ME6229