Tags

Type your tag names separated by a space and hit enter

Initiation of DNA replication in a ColE1-type plasmid: isolation of mutations in the ori region.
Proc Natl Acad Sci U S A. 1980 Nov; 77(11):6744-8.PN

Abstract

We have constructed a plaque-forming hybrid phage, lambda SN4, which behaves as a composite replicon of the lambda phage and a mini-ColE1 plasmid. From the hybrid phage, plaque-type mutants altered in the ability to replicate as a ColE1 replicon were isolated. These mutations were designated as cer, signifying ColE1 replication defective. One of such mutants, lambda SN4cer6, was studied further. The mutant DNA was unable to replicate in vivo if expression and function of its lambda replicon were inhibited. The defect could not be complemented in trans. DNA sequence determination of the mutant phage revealed a single base pair (bp) alteration, C-G to T-A, at 160 bp upstream from the ori site of its ColE1 replicon. From lambda SN4cer6, revertants were obtained that had regained function of the ColE1 replicon, and they could be classified into two groups that showed a full and a partial recovery in the rates of ColE1-driven DNA synthesis. DNA sequence determination of revertant DNA indicated that the former group contained true revertants, T-A to C-G, at the cer6 site, whereas one of the partial revertants was found to sustain a secondary-site mutation, G-C to A-T at 187 bp upstream of the ori site. It was possible to construct a hairpin structure that starts by hydrogen bonding of bases at the site -160 and -187.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

6256758

Citation

Naito, S, and H Uchida. "Initiation of DNA Replication in a ColE1-type Plasmid: Isolation of Mutations in the Ori Region." Proceedings of the National Academy of Sciences of the United States of America, vol. 77, no. 11, 1980, pp. 6744-8.
Naito S, Uchida H. Initiation of DNA replication in a ColE1-type plasmid: isolation of mutations in the ori region. Proc Natl Acad Sci USA. 1980;77(11):6744-8.
Naito, S., & Uchida, H. (1980). Initiation of DNA replication in a ColE1-type plasmid: isolation of mutations in the ori region. Proceedings of the National Academy of Sciences of the United States of America, 77(11), 6744-8.
Naito S, Uchida H. Initiation of DNA Replication in a ColE1-type Plasmid: Isolation of Mutations in the Ori Region. Proc Natl Acad Sci USA. 1980;77(11):6744-8. PubMed PMID: 6256758.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Initiation of DNA replication in a ColE1-type plasmid: isolation of mutations in the ori region. AU - Naito,S, AU - Uchida,H, PY - 1980/11/1/pubmed PY - 1980/11/1/medline PY - 1980/11/1/entrez SP - 6744 EP - 8 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc. Natl. Acad. Sci. U.S.A. VL - 77 IS - 11 N2 - We have constructed a plaque-forming hybrid phage, lambda SN4, which behaves as a composite replicon of the lambda phage and a mini-ColE1 plasmid. From the hybrid phage, plaque-type mutants altered in the ability to replicate as a ColE1 replicon were isolated. These mutations were designated as cer, signifying ColE1 replication defective. One of such mutants, lambda SN4cer6, was studied further. The mutant DNA was unable to replicate in vivo if expression and function of its lambda replicon were inhibited. The defect could not be complemented in trans. DNA sequence determination of the mutant phage revealed a single base pair (bp) alteration, C-G to T-A, at 160 bp upstream from the ori site of its ColE1 replicon. From lambda SN4cer6, revertants were obtained that had regained function of the ColE1 replicon, and they could be classified into two groups that showed a full and a partial recovery in the rates of ColE1-driven DNA synthesis. DNA sequence determination of revertant DNA indicated that the former group contained true revertants, T-A to C-G, at the cer6 site, whereas one of the partial revertants was found to sustain a secondary-site mutation, G-C to A-T at 187 bp upstream of the ori site. It was possible to construct a hairpin structure that starts by hydrogen bonding of bases at the site -160 and -187. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/6256758/Initiation_of_DNA_replication_in_a_ColE1_type_plasmid:_isolation_of_mutations_in_the_ori_region_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/6256758/ DB - PRIME DP - Unbound Medicine ER -