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Isolation and characterization of ColE1-derived plasmid copy-number mutant.
Proc Natl Acad Sci U S A. 1978 Dec; 75(12):5869-73.PN

Abstract

The plasmid pBGP120 is a ColE1 derivative that contains elements of the Escherichia coli lac operon and the Tn3 transposon. We have selected and isolated a copy-number mutant of pBGP120. In exponentially growing cultures, the copy-number mutant, pOP1, represents approximately 30% of total intracellular DNA compared to about 5% for pBGP120. Plasmid-encoded beta-galactosidase monomer can represent 50% of newly synthesized protein in cells carrying pOP1. pOP1 is structurally unstable in certain genetic backgrounds and under certain growth conditions, breaking down to a smaller sized plasmid that retains the DNA overproducer phenotype and the Tn3 transposon. The smaller overproducer plasmid, pOP1delta6, is generated by a continuous deletion of sequences located between one end of the Tn3 transposon and a site about 630 nucleotides from the EcoRI site in the beta-galactosidase structural gene of pOP1. pOP1delta6 retains the ColE1 origin of replication but has lost the lac promotor and operator and most of the beta-galactosidase structural gene. pOP1delta6 exists at approximately 210 copies per chromosome in exponentially growing cells.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

104293

Citation

Gelfand, D H., et al. "Isolation and Characterization of ColE1-derived Plasmid Copy-number Mutant." Proceedings of the National Academy of Sciences of the United States of America, vol. 75, no. 12, 1978, pp. 5869-73.
Gelfand DH, Shepard HM, O'Farrell PH, et al. Isolation and characterization of ColE1-derived plasmid copy-number mutant. Proc Natl Acad Sci USA. 1978;75(12):5869-73.
Gelfand, D. H., Shepard, H. M., O'Farrell, P. H., & Polisky, B. (1978). Isolation and characterization of ColE1-derived plasmid copy-number mutant. Proceedings of the National Academy of Sciences of the United States of America, 75(12), 5869-73.
Gelfand DH, et al. Isolation and Characterization of ColE1-derived Plasmid Copy-number Mutant. Proc Natl Acad Sci USA. 1978;75(12):5869-73. PubMed PMID: 104293.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and characterization of ColE1-derived plasmid copy-number mutant. AU - Gelfand,D H, AU - Shepard,H M, AU - O'Farrell,P H, AU - Polisky,B, PY - 1978/12/1/pubmed PY - 1978/12/1/medline PY - 1978/12/1/entrez SP - 5869 EP - 73 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc. Natl. Acad. Sci. U.S.A. VL - 75 IS - 12 N2 - The plasmid pBGP120 is a ColE1 derivative that contains elements of the Escherichia coli lac operon and the Tn3 transposon. We have selected and isolated a copy-number mutant of pBGP120. In exponentially growing cultures, the copy-number mutant, pOP1, represents approximately 30% of total intracellular DNA compared to about 5% for pBGP120. Plasmid-encoded beta-galactosidase monomer can represent 50% of newly synthesized protein in cells carrying pOP1. pOP1 is structurally unstable in certain genetic backgrounds and under certain growth conditions, breaking down to a smaller sized plasmid that retains the DNA overproducer phenotype and the Tn3 transposon. The smaller overproducer plasmid, pOP1delta6, is generated by a continuous deletion of sequences located between one end of the Tn3 transposon and a site about 630 nucleotides from the EcoRI site in the beta-galactosidase structural gene of pOP1. pOP1delta6 retains the ColE1 origin of replication but has lost the lac promotor and operator and most of the beta-galactosidase structural gene. pOP1delta6 exists at approximately 210 copies per chromosome in exponentially growing cells. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/104293/Isolation_and_characterization_of_ColE1_derived_plasmid_copy_number_mutant_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/104293/ DB - PRIME DP - Unbound Medicine ER -