Tags

Type your tag names separated by a space and hit enter

Characterization of pEC156, a ColE1-type plasmid from Escherichia coli E1585-68 that carries genes of the EcoVIII restriction-modification system.
Plasmid 2001; 46(2):128-39P

Abstract

The complete 4312-bp sequence of the pEC156 plasmid from Escherichia coli E1585-68, which carries genes encoding the EcoVIII restriction-modification (R-M) system, an isoschizomer of HindIII from Haemophilus influenzae, has been determined. Two clustered and convergently oriented open reading frames, large enough to encode genes of the EcoVIII R-M system, were found. The transcriptional start points were mapped by the primer extension method. The relative molecular masses of the EcoVIII endonuclease and EcoVIII methyltransferase deduced from the nucleotide sequence are 35,554 and 33,910, respectively. Nucleotide sequence analysis of pEC156 suggests that this plasmid is a ColE1-type replicon. It consists of an origin of replication and two untranslated genes encoding RNA I and RNA II, both involved in the regulation of plasmid DNA replication. The replication region also contains the gene encoding a 64-aa Rom-like protein. Inactivation of the putative rom gene by insertion of a kanamycin-resistance cassette resulted in 4.5-fold increase in pEC156-derived plasmid copy number in E. coli cells. All of these elements (RNA I, RNA II, and rom) reveal a high level of similarity to ColE1 homologs. The replication of all ColE1-type plasmids is dependent on the activity of E. coli DNA polymerase I. It was shown that a pEC156 derivative (pIB8) carrying an antibiotic resistance gene indeed failed to replicate in an E. coli polA12(ts) mutant at 43 degrees C, and its copy number was reduced in the E. coli pcnB80 mutant. These results prove that pEC156 is a ColE1-type replicon.

Authors+Show Affiliations

Department of Microbiology, University of Gdańsk, Kładki 24, Gdańsk, 80-822, Poland.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

11591138

Citation

Mruk, I, et al. "Characterization of pEC156, a ColE1-type Plasmid From Escherichia Coli E1585-68 That Carries Genes of the EcoVIII Restriction-modification System." Plasmid, vol. 46, no. 2, 2001, pp. 128-39.
Mruk I, Sektas M, Kaczorowski T. Characterization of pEC156, a ColE1-type plasmid from Escherichia coli E1585-68 that carries genes of the EcoVIII restriction-modification system. Plasmid. 2001;46(2):128-39.
Mruk, I., Sektas, M., & Kaczorowski, T. (2001). Characterization of pEC156, a ColE1-type plasmid from Escherichia coli E1585-68 that carries genes of the EcoVIII restriction-modification system. Plasmid, 46(2), pp. 128-39.
Mruk I, Sektas M, Kaczorowski T. Characterization of pEC156, a ColE1-type Plasmid From Escherichia Coli E1585-68 That Carries Genes of the EcoVIII Restriction-modification System. Plasmid. 2001;46(2):128-39. PubMed PMID: 11591138.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of pEC156, a ColE1-type plasmid from Escherichia coli E1585-68 that carries genes of the EcoVIII restriction-modification system. AU - Mruk,I, AU - Sektas,M, AU - Kaczorowski,T, PY - 2001/10/10/pubmed PY - 2002/1/5/medline PY - 2001/10/10/entrez SP - 128 EP - 39 JF - Plasmid JO - Plasmid VL - 46 IS - 2 N2 - The complete 4312-bp sequence of the pEC156 plasmid from Escherichia coli E1585-68, which carries genes encoding the EcoVIII restriction-modification (R-M) system, an isoschizomer of HindIII from Haemophilus influenzae, has been determined. Two clustered and convergently oriented open reading frames, large enough to encode genes of the EcoVIII R-M system, were found. The transcriptional start points were mapped by the primer extension method. The relative molecular masses of the EcoVIII endonuclease and EcoVIII methyltransferase deduced from the nucleotide sequence are 35,554 and 33,910, respectively. Nucleotide sequence analysis of pEC156 suggests that this plasmid is a ColE1-type replicon. It consists of an origin of replication and two untranslated genes encoding RNA I and RNA II, both involved in the regulation of plasmid DNA replication. The replication region also contains the gene encoding a 64-aa Rom-like protein. Inactivation of the putative rom gene by insertion of a kanamycin-resistance cassette resulted in 4.5-fold increase in pEC156-derived plasmid copy number in E. coli cells. All of these elements (RNA I, RNA II, and rom) reveal a high level of similarity to ColE1 homologs. The replication of all ColE1-type plasmids is dependent on the activity of E. coli DNA polymerase I. It was shown that a pEC156 derivative (pIB8) carrying an antibiotic resistance gene indeed failed to replicate in an E. coli polA12(ts) mutant at 43 degrees C, and its copy number was reduced in the E. coli pcnB80 mutant. These results prove that pEC156 is a ColE1-type replicon. SN - 0147-619X UR - https://www.unboundmedicine.com/medline/citation/11591138/Characterization_of_pEC156_a_ColE1_type_plasmid_from_Escherichia_coli_E1585_68_that_carries_genes_of_the_EcoVIII_restriction_modification_system_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0147-619X(01)91534-3 DB - PRIME DP - Unbound Medicine ER -