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Characterization of pMRI 5.2, a rolling-circle-type plasmid from Lactobacillus plantarum BFE 5092 which harbours two different replication initiation genes.
Plasmid 2013; 69(2):160-71P

Abstract

Plasmid pMRI 5.2 from Lactobacillus plantarum BFE 5092 was sequenced and analysed. The sequence consists of 5206bp with a mol% G+C content of 35.8%. Nine putative open reading frames were identified. A typical pC194 family double strand origin (dso) and a putative single strand origin (sso) were predicted upstream of a rep gene. This rep gene encoded a replication protein of 314 amino acids exhibiting 98% amino acid sequence identity to the Rep protein of plasmid pLAB1000 from Lactobacillus hilgardii. A mob gene encoding a mobilization protein was also identified and this protein showed high amino acid similarity to Mob proteins from various L. plantarum plasmids. Downstream of the mob gene, a second putative replication region was identified that is similar to the pMV158 family of plasmids. It contains a dso as well as a putative sso, and encodes the 52 amino acid repressor-like protein RepA, the replication initiation protein RepB of 215 amino acids, and the 48 amino acid RepC that is similar to ORFD of the lactococcal plasmid pWVO1. RT-PCR and qRT-PCR expression analyses of the rep and repB genes showed that the repB gene was expressed at a higher level. To confirm that the plasmid replicated by the rolling-circle-type mechanism, the presence of a characteristic single strand intermediate DNA was shown to be produced during replication. Plasmid copy number was ca. 30 per equivalent chromosome copy number based on qRT-PCR analyses. The plasmid also encodes four additional putative proteins of unknown function. The unusual feature of a rolling-circle plasmid having two different plasmid-encoded replication initiation proteins from different replicon families suggests that the genes for these may have originated from different plasmids.

Authors+Show Affiliations

Department of Safety and Quality of Fruit and Vegetables, Max Rubner-Institute, Federal Research Institute for Nutrition and Food, Haid-und-Neu-Strasse 9, D-76131 Karlsruhe, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

23220639

Citation

Cho, Gyu-Sung, et al. "Characterization of pMRI 5.2, a Rolling-circle-type Plasmid From Lactobacillus Plantarum BFE 5092 Which Harbours Two Different Replication Initiation Genes." Plasmid, vol. 69, no. 2, 2013, pp. 160-71.
Cho GS, Huch M, Mathara JM, et al. Characterization of pMRI 5.2, a rolling-circle-type plasmid from Lactobacillus plantarum BFE 5092 which harbours two different replication initiation genes. Plasmid. 2013;69(2):160-71.
Cho, G. S., Huch, M., Mathara, J. M., van Belkum, M. J., & Franz, C. M. (2013). Characterization of pMRI 5.2, a rolling-circle-type plasmid from Lactobacillus plantarum BFE 5092 which harbours two different replication initiation genes. Plasmid, 69(2), pp. 160-71. doi:10.1016/j.plasmid.2012.11.005.
Cho GS, et al. Characterization of pMRI 5.2, a Rolling-circle-type Plasmid From Lactobacillus Plantarum BFE 5092 Which Harbours Two Different Replication Initiation Genes. Plasmid. 2013;69(2):160-71. PubMed PMID: 23220639.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of pMRI 5.2, a rolling-circle-type plasmid from Lactobacillus plantarum BFE 5092 which harbours two different replication initiation genes. AU - Cho,Gyu-Sung, AU - Huch,Melanie, AU - Mathara,Julius Maina, AU - van Belkum,Marco J, AU - Franz,Charles M A P, Y1 - 2012/12/06/ PY - 2012/07/26/received PY - 2012/11/28/revised PY - 2012/11/29/accepted PY - 2012/12/11/entrez PY - 2012/12/12/pubmed PY - 2014/1/11/medline SP - 160 EP - 71 JF - Plasmid JO - Plasmid VL - 69 IS - 2 N2 - Plasmid pMRI 5.2 from Lactobacillus plantarum BFE 5092 was sequenced and analysed. The sequence consists of 5206bp with a mol% G+C content of 35.8%. Nine putative open reading frames were identified. A typical pC194 family double strand origin (dso) and a putative single strand origin (sso) were predicted upstream of a rep gene. This rep gene encoded a replication protein of 314 amino acids exhibiting 98% amino acid sequence identity to the Rep protein of plasmid pLAB1000 from Lactobacillus hilgardii. A mob gene encoding a mobilization protein was also identified and this protein showed high amino acid similarity to Mob proteins from various L. plantarum plasmids. Downstream of the mob gene, a second putative replication region was identified that is similar to the pMV158 family of plasmids. It contains a dso as well as a putative sso, and encodes the 52 amino acid repressor-like protein RepA, the replication initiation protein RepB of 215 amino acids, and the 48 amino acid RepC that is similar to ORFD of the lactococcal plasmid pWVO1. RT-PCR and qRT-PCR expression analyses of the rep and repB genes showed that the repB gene was expressed at a higher level. To confirm that the plasmid replicated by the rolling-circle-type mechanism, the presence of a characteristic single strand intermediate DNA was shown to be produced during replication. Plasmid copy number was ca. 30 per equivalent chromosome copy number based on qRT-PCR analyses. The plasmid also encodes four additional putative proteins of unknown function. The unusual feature of a rolling-circle plasmid having two different plasmid-encoded replication initiation proteins from different replicon families suggests that the genes for these may have originated from different plasmids. SN - 1095-9890 UR - https://www.unboundmedicine.com/medline/citation/23220639/Characterization_of_pMRI_5_2_a_rolling_circle_type_plasmid_from_Lactobacillus_plantarum_BFE_5092_which_harbours_two_different_replication_initiation_genes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0147-619X(12)00112-6 DB - PRIME DP - Unbound Medicine ER -