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Characterization of the theta replication plasmid pGR7 from Acetobacter aceti CCM 3610.
Res Microbiol. 2012 Jul; 163(6-7):419-26.RM

Abstract

A cryptic plasmid of Acetobacter aceti CCM 3610, designated pGR7, was sequenced and characterized. It is a 2446-bp circular molecule with a G + C content of 30%, which is unusual when compared to the already known plasmids isolated from Acetobacter genera. Sequence analysis of pGR7 revealed three putative open reading frames (ORFs). ORF1 displays low similarity with other Acetobacter plasmid replication proteins. The other two ORFs show similarities only to hypothetical proteins and do not encode any important protein. The replication module comprises a DnaA box-like sequence, indirect repeats, a potential prokaryotic promoter and the rep gene. The rep module organization is similar to that found in other theta-replicating plasmids from acetic acid bacteria that stably maintain in both Acetobacter and Escherichia coli, with two repeated sequences containing modules. Nevertheless, the pGR7 plasmid could replicate and be stably maintained only in Acetobacter strains and not in E. coli, another uncommon feature of this plasmid. The Rep protein was cloned into the pET30a + expression vector and purified by high-performance liquid chromatography. The helicase activity was determined and the ability of the protein to bind to the plasmid regulation region was confirmed by an electrophoretic mobility shift assay. The plasmid was stable in the Acetobacter cells after cultivation under nonselective conditions. By real-time polymerase chain reaction, the relative copy number of pGR7 was estimated to be seven copies per host chromosome equivalent.

Authors+Show Affiliations

Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Gent, Belgium. pegro@psb.vib-ugent.beNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

22842078

Citation

Grones, Peter, and Jozef Grones. "Characterization of the Theta Replication Plasmid pGR7 From Acetobacter Aceti CCM 3610." Research in Microbiology, vol. 163, no. 6-7, 2012, pp. 419-26.
Grones P, Grones J. Characterization of the theta replication plasmid pGR7 from Acetobacter aceti CCM 3610. Res Microbiol. 2012;163(6-7):419-26.
Grones, P., & Grones, J. (2012). Characterization of the theta replication plasmid pGR7 from Acetobacter aceti CCM 3610. Research in Microbiology, 163(6-7), 419-26. https://doi.org/10.1016/j.resmic.2012.07.002
Grones P, Grones J. Characterization of the Theta Replication Plasmid pGR7 From Acetobacter Aceti CCM 3610. Res Microbiol. 2012;163(6-7):419-26. PubMed PMID: 22842078.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of the theta replication plasmid pGR7 from Acetobacter aceti CCM 3610. AU - Grones,Peter, AU - Grones,Jozef, Y1 - 2012/07/24/ PY - 2012/02/15/received PY - 2012/06/21/accepted PY - 2012/7/31/entrez PY - 2012/7/31/pubmed PY - 2013/1/17/medline SP - 419 EP - 26 JF - Research in microbiology JO - Res Microbiol VL - 163 IS - 6-7 N2 - A cryptic plasmid of Acetobacter aceti CCM 3610, designated pGR7, was sequenced and characterized. It is a 2446-bp circular molecule with a G + C content of 30%, which is unusual when compared to the already known plasmids isolated from Acetobacter genera. Sequence analysis of pGR7 revealed three putative open reading frames (ORFs). ORF1 displays low similarity with other Acetobacter plasmid replication proteins. The other two ORFs show similarities only to hypothetical proteins and do not encode any important protein. The replication module comprises a DnaA box-like sequence, indirect repeats, a potential prokaryotic promoter and the rep gene. The rep module organization is similar to that found in other theta-replicating plasmids from acetic acid bacteria that stably maintain in both Acetobacter and Escherichia coli, with two repeated sequences containing modules. Nevertheless, the pGR7 plasmid could replicate and be stably maintained only in Acetobacter strains and not in E. coli, another uncommon feature of this plasmid. The Rep protein was cloned into the pET30a + expression vector and purified by high-performance liquid chromatography. The helicase activity was determined and the ability of the protein to bind to the plasmid regulation region was confirmed by an electrophoretic mobility shift assay. The plasmid was stable in the Acetobacter cells after cultivation under nonselective conditions. By real-time polymerase chain reaction, the relative copy number of pGR7 was estimated to be seven copies per host chromosome equivalent. SN - 1769-7123 UR - https://www.unboundmedicine.com/medline/citation/22842078/Characterization_of_the_theta_replication_plasmid_pGR7_from_Acetobacter_aceti_CCM_3610_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923-2508(12)00097-6 DB - PRIME DP - Unbound Medicine ER -