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Transcription of the replication control region of the IncFII R-plasmid NR1 in vitro and in vivo.
J Mol Biol. 1985 Feb 05; 181(3):395-410.JM

Abstract

The minimal replicon of the 90,000 base-pair IncFII R plasmid NR1 consists of a 2700 base-pair region of the DNA. Minireplicator plasmids consisting of the 2700 base-pair minimal replicon plus a 2200 base-pair region coding for chloramphenicol acetyltransferase (cat) were used as templates for in vitro transcription. Six RNA transcripts were synthesized from these templates in vitro. We have determined the directions of transcription and the approximate sites of initiation and termination of each of the in vitro RNA transcripts. One RNA transcript was synthesized from the cat gene, while the other five were transcribed from the minimal replicon. Four of the RNA transcripts also were identified by quantitative hybridization of RNA synthesized in vivo from these minireplicator plasmids. The strengths of the promoters for the RNA transcripts were estimated by the relative rates of transcription both in vitro and in vivo. Transcription from convergent promoters reduced the rate of RNA synthesis in vivo in both directions. In vivo, a significant fraction of the cat mRNA was extended past its in vitro termination point. Transcription of mutants that have altered plasmid copy number and/or incompatibility properties also were examined. The possible roles of each of the transcripts as mRNA and their involvement in regulation of DNA replication are discussed.

Authors

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

Pub Type(s)

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

Language

eng

PubMed ID

2580099

Citation

Womble, D D., et al. "Transcription of the Replication Control Region of the IncFII R-plasmid NR1 in Vitro and in Vivo." Journal of Molecular Biology, vol. 181, no. 3, 1985, pp. 395-410.
Womble DD, Sampathkumar P, Easton AM, et al. Transcription of the replication control region of the IncFII R-plasmid NR1 in vitro and in vivo. J Mol Biol. 1985;181(3):395-410.
Womble, D. D., Sampathkumar, P., Easton, A. M., Luckow, V. A., & Rownd, R. H. (1985). Transcription of the replication control region of the IncFII R-plasmid NR1 in vitro and in vivo. Journal of Molecular Biology, 181(3), 395-410.
Womble DD, et al. Transcription of the Replication Control Region of the IncFII R-plasmid NR1 in Vitro and in Vivo. J Mol Biol. 1985 Feb 5;181(3):395-410. PubMed PMID: 2580099.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transcription of the replication control region of the IncFII R-plasmid NR1 in vitro and in vivo. AU - Womble,D D, AU - Sampathkumar,P, AU - Easton,A M, AU - Luckow,V A, AU - Rownd,R H, PY - 1985/2/5/pubmed PY - 1985/2/5/medline PY - 1985/2/5/entrez SP - 395 EP - 410 JF - Journal of molecular biology JO - J. Mol. Biol. VL - 181 IS - 3 N2 - The minimal replicon of the 90,000 base-pair IncFII R plasmid NR1 consists of a 2700 base-pair region of the DNA. Minireplicator plasmids consisting of the 2700 base-pair minimal replicon plus a 2200 base-pair region coding for chloramphenicol acetyltransferase (cat) were used as templates for in vitro transcription. Six RNA transcripts were synthesized from these templates in vitro. We have determined the directions of transcription and the approximate sites of initiation and termination of each of the in vitro RNA transcripts. One RNA transcript was synthesized from the cat gene, while the other five were transcribed from the minimal replicon. Four of the RNA transcripts also were identified by quantitative hybridization of RNA synthesized in vivo from these minireplicator plasmids. The strengths of the promoters for the RNA transcripts were estimated by the relative rates of transcription both in vitro and in vivo. Transcription from convergent promoters reduced the rate of RNA synthesis in vivo in both directions. In vivo, a significant fraction of the cat mRNA was extended past its in vitro termination point. Transcription of mutants that have altered plasmid copy number and/or incompatibility properties also were examined. The possible roles of each of the transcripts as mRNA and their involvement in regulation of DNA replication are discussed. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/2580099/Transcription_of_the_replication_control_region_of_the_IncFII_R_plasmid_NR1_in_vitro_and_in_vivo_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0022-2836(85)90228-1 DB - PRIME DP - Unbound Medicine ER -