Nucleotide sequence and analysis of pRC12 and pRC18, two theta-replicating plasmids harbored by Lactobacillus curvatus CRL 705
The nucleotide sequences of plasmids pRC12 (12,342 bp; GC 43.99%) and pRC18 (18,664 bp; GC 34.33%), harbored by the bacteriocin-producer Lactobacillus curvatus CRL 705, were determined and analyzed. Plasmids pRC12 and pRC18 share a region with high DNA identity (> 83% identity between RepA, a Typ...
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| Veröffentlicht in: | PloS one 2020-04, Vol.15 (4), p.e0230857-e0230857 |
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| creator | Terán, Lucrecia C Cuozzo, Sergio A Aristimuño Ficoseco, María C Fadda, Silvina Chaillou, Stéphane Champomier-Vergès, Marie-Christine Zagorec, Monique Hébert, Elvira M Raya, Raúl R |
| description | The nucleotide sequences of plasmids pRC12 (12,342 bp; GC 43.99%) and pRC18 (18,664 bp; GC 34.33%), harbored by the bacteriocin-producer Lactobacillus curvatus CRL 705, were determined and analyzed. Plasmids pRC12 and pRC18 share a region with high DNA identity (> 83% identity between RepA, a Type II toxin-antitoxin system and a tyrosine integrase genes) and are stably maintained in their natural host L. curvatus CRL 705. Both plasmids are low copy number and belong to the theta-type replicating group. While pRC12 is a pUCL287-like plasmid that possesses iterons and the repA and repB genes for replication, pRC18 harbors a 168 amino acid replication protein affiliated to RepB, which was named RepB'. Plasmid pRC18 also possesses a pUCL287-like repA gene but it was disrupted by an 11 kb insertion element that contains RepB', several transposases/IS elements, and the lactocin Lac705 operon. An Escherichia coli / Lactobacillus shuttle vector, named plasmid p3B1, carrying the pRC18 replicon (i.e. repB' and replication origin), a chloramphenicol resistance gene and a pBluescript backbone, was constructed and used to define the host range of RepB'. Chloramphenicol-resistant transformants were obtained after electroporation of Lactobacillus plantarum CRL 691, Lactobacillus sakei 23K and a plasmid-cured derivative of L. curvatus CRL 705, but not of L. curvatus DSM 20019 or Lactococcus lactis NZ9000. Depending on the host, transformation efficiency ranged from 102 to 107 per μg of DNA; in the new hosts, the plasmid was relatively stable as 29-53% of recombinants kept it after cell growth for 100 generations in the absence of selective pressure. Plasmid p3B1 could therefore be used for cloning and functional studies in several Lactobacillus species. |
| doi_str_mv | 10.1371/journal.pone.0230857 |
| format | Article |
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Plasmids pRC12 and pRC18 share a region with high DNA identity (> 83% identity between RepA, a Type II toxin-antitoxin system and a tyrosine integrase genes) and are stably maintained in their natural host L. curvatus CRL 705. Both plasmids are low copy number and belong to the theta-type replicating group. While pRC12 is a pUCL287-like plasmid that possesses iterons and the repA and repB genes for replication, pRC18 harbors a 168 amino acid replication protein affiliated to RepB, which was named RepB'. Plasmid pRC18 also possesses a pUCL287-like repA gene but it was disrupted by an 11 kb insertion element that contains RepB', several transposases/IS elements, and the lactocin Lac705 operon. An Escherichia coli / Lactobacillus shuttle vector, named plasmid p3B1, carrying the pRC18 replicon (i.e. repB' and replication origin), a chloramphenicol resistance gene and a pBluescript backbone, was constructed and used to define the host range of RepB'. Chloramphenicol-resistant transformants were obtained after electroporation of Lactobacillus plantarum CRL 691, Lactobacillus sakei 23K and a plasmid-cured derivative of L. curvatus CRL 705, but not of L. curvatus DSM 20019 or Lactococcus lactis NZ9000. Depending on the host, transformation efficiency ranged from 102 to 107 per μg of DNA; in the new hosts, the plasmid was relatively stable as 29-53% of recombinants kept it after cell growth for 100 generations in the absence of selective pressure. Plasmid p3B1 could therefore be used for cloning and functional studies in several Lactobacillus species.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0230857</identifier><identifier>PMID: 32240216</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amino Acid Sequence - genetics ; Amino acids ; Analysis ; Antibacterial agents ; Antitoxins ; Bacteria ; Bacterial Proteins - genetics ; Bacteriocins ; Base sequence ; Base Sequence - genetics ; Biological products ; Biology and life sciences ; Chloramphenicol ; Chloromycetin ; Chromosomes ; Cloning ; Copy number ; Deoxyribonucleic acid ; DNA ; DNA Replication - genetics ; DNA, Bacterial - genetics ; E coli ; Electroporation ; Gene expression ; Genes ; Genetic engineering ; Genetic vectors ; Genetic Vectors - genetics ; Genomes ; Host range ; Identity ; Integrase ; Lactobacillus ; Lactobacillus - genetics ; Lactobacillus curvatus ; Life Sciences ; Medicine and Health Sciences ; Nucleotide sequence ; Nucleotides ; Phenols (Class of compounds) ; Plasmids ; Plasmids - genetics ; Recombinants ; RepA gene ; Replicating ; Replication ; Replication Origin - genetics ; Replication origins ; Replicon - genetics ; Research and analysis methods ; Sequence Analysis, DNA - methods ; Toxins ; Transposases - genetics ; Tyrosine ; Vectors (Biology)</subject><ispartof>PloS one, 2020-04, Vol.15 (4), p.e0230857-e0230857</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Terán et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Attribution</rights><rights>2020 Terán et al 2020 Terán et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c726t-e470412930b1915a56e22678d4ea164515211367b53652892e3791f41e5fb9e33</citedby><cites>FETCH-LOGICAL-c726t-e470412930b1915a56e22678d4ea164515211367b53652892e3791f41e5fb9e33</cites><orcidid>0000-0002-6387-160X ; 0000-0003-2229-0697 ; 0000-0001-7320-538X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117683/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117683/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32240216$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.inrae.fr/hal-02921909$$DView record in HAL$$Hfree_for_read</backlink></links><search><contributor>Singh, Anirudh K.</contributor><creatorcontrib>Terán, Lucrecia C</creatorcontrib><creatorcontrib>Cuozzo, Sergio A</creatorcontrib><creatorcontrib>Aristimuño Ficoseco, María C</creatorcontrib><creatorcontrib>Fadda, Silvina</creatorcontrib><creatorcontrib>Chaillou, Stéphane</creatorcontrib><creatorcontrib>Champomier-Vergès, Marie-Christine</creatorcontrib><creatorcontrib>Zagorec, Monique</creatorcontrib><creatorcontrib>Hébert, Elvira M</creatorcontrib><creatorcontrib>Raya, Raúl R</creatorcontrib><title>Nucleotide sequence and analysis of pRC12 and pRC18, two theta-replicating plasmids harbored by Lactobacillus curvatus CRL 705</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The nucleotide sequences of plasmids pRC12 (12,342 bp; GC 43.99%) and pRC18 (18,664 bp; GC 34.33%), harbored by the bacteriocin-producer Lactobacillus curvatus CRL 705, were determined and analyzed. Plasmids pRC12 and pRC18 share a region with high DNA identity (> 83% identity between RepA, a Type II toxin-antitoxin system and a tyrosine integrase genes) and are stably maintained in their natural host L. curvatus CRL 705. Both plasmids are low copy number and belong to the theta-type replicating group. While pRC12 is a pUCL287-like plasmid that possesses iterons and the repA and repB genes for replication, pRC18 harbors a 168 amino acid replication protein affiliated to RepB, which was named RepB'. Plasmid pRC18 also possesses a pUCL287-like repA gene but it was disrupted by an 11 kb insertion element that contains RepB', several transposases/IS elements, and the lactocin Lac705 operon. An Escherichia coli / Lactobacillus shuttle vector, named plasmid p3B1, carrying the pRC18 replicon (i.e. repB' and replication origin), a chloramphenicol resistance gene and a pBluescript backbone, was constructed and used to define the host range of RepB'. Chloramphenicol-resistant transformants were obtained after electroporation of Lactobacillus plantarum CRL 691, Lactobacillus sakei 23K and a plasmid-cured derivative of L. curvatus CRL 705, but not of L. curvatus DSM 20019 or Lactococcus lactis NZ9000. Depending on the host, transformation efficiency ranged from 102 to 107 per μg of DNA; in the new hosts, the plasmid was relatively stable as 29-53% of recombinants kept it after cell growth for 100 generations in the absence of selective pressure. Plasmid p3B1 could therefore be used for cloning and functional studies in several Lactobacillus species.</description><subject>Amino Acid Sequence - genetics</subject><subject>Amino acids</subject><subject>Analysis</subject><subject>Antibacterial agents</subject><subject>Antitoxins</subject><subject>Bacteria</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacteriocins</subject><subject>Base sequence</subject><subject>Base Sequence - genetics</subject><subject>Biological products</subject><subject>Biology and life sciences</subject><subject>Chloramphenicol</subject><subject>Chloromycetin</subject><subject>Chromosomes</subject><subject>Cloning</subject><subject>Copy number</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Replication - genetics</subject><subject>DNA, Bacterial - genetics</subject><subject>E coli</subject><subject>Electroporation</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Genetic engineering</subject><subject>Genetic vectors</subject><subject>Genetic Vectors - genetics</subject><subject>Genomes</subject><subject>Host range</subject><subject>Identity</subject><subject>Integrase</subject><subject>Lactobacillus</subject><subject>Lactobacillus - genetics</subject><subject>Lactobacillus curvatus</subject><subject>Life Sciences</subject><subject>Medicine and Health Sciences</subject><subject>Nucleotide sequence</subject><subject>Nucleotides</subject><subject>Phenols (Class of compounds)</subject><subject>Plasmids</subject><subject>Plasmids - genetics</subject><subject>Recombinants</subject><subject>RepA gene</subject><subject>Replicating</subject><subject>Replication</subject><subject>Replication Origin - genetics</subject><subject>Replication origins</subject><subject>Replicon - genetics</subject><subject>Research and analysis methods</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Toxins</subject><subject>Transposases - genetics</subject><subject>Tyrosine</subject><subject>Vectors 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Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Terán, Lucrecia C</au><au>Cuozzo, Sergio A</au><au>Aristimuño Ficoseco, María C</au><au>Fadda, Silvina</au><au>Chaillou, Stéphane</au><au>Champomier-Vergès, Marie-Christine</au><au>Zagorec, Monique</au><au>Hébert, Elvira M</au><au>Raya, Raúl R</au><au>Singh, Anirudh K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nucleotide sequence and analysis of pRC12 and pRC18, two theta-replicating plasmids harbored by Lactobacillus curvatus CRL 705</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2020-04-02</date><risdate>2020</risdate><volume>15</volume><issue>4</issue><spage>e0230857</spage><epage>e0230857</epage><pages>e0230857-e0230857</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>The nucleotide sequences of plasmids pRC12 (12,342 bp; GC 43.99%) and pRC18 (18,664 bp; GC 34.33%), harbored by the bacteriocin-producer Lactobacillus curvatus CRL 705, were determined and analyzed. Plasmids pRC12 and pRC18 share a region with high DNA identity (> 83% identity between RepA, a Type II toxin-antitoxin system and a tyrosine integrase genes) and are stably maintained in their natural host L. curvatus CRL 705. Both plasmids are low copy number and belong to the theta-type replicating group. While pRC12 is a pUCL287-like plasmid that possesses iterons and the repA and repB genes for replication, pRC18 harbors a 168 amino acid replication protein affiliated to RepB, which was named RepB'. Plasmid pRC18 also possesses a pUCL287-like repA gene but it was disrupted by an 11 kb insertion element that contains RepB', several transposases/IS elements, and the lactocin Lac705 operon. An Escherichia coli / Lactobacillus shuttle vector, named plasmid p3B1, carrying the pRC18 replicon (i.e. repB' and replication origin), a chloramphenicol resistance gene and a pBluescript backbone, was constructed and used to define the host range of RepB'. Chloramphenicol-resistant transformants were obtained after electroporation of Lactobacillus plantarum CRL 691, Lactobacillus sakei 23K and a plasmid-cured derivative of L. curvatus CRL 705, but not of L. curvatus DSM 20019 or Lactococcus lactis NZ9000. Depending on the host, transformation efficiency ranged from 102 to 107 per μg of DNA; in the new hosts, the plasmid was relatively stable as 29-53% of recombinants kept it after cell growth for 100 generations in the absence of selective pressure. Plasmid p3B1 could therefore be used for cloning and functional studies in several Lactobacillus species.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>32240216</pmid><doi>10.1371/journal.pone.0230857</doi><tpages>e0230857</tpages><orcidid>https://orcid.org/0000-0002-6387-160X</orcidid><orcidid>https://orcid.org/0000-0003-2229-0697</orcidid><orcidid>https://orcid.org/0000-0001-7320-538X</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2020-04, Vol.15 (4), p.e0230857-e0230857 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2385711148 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence - genetics Amino acids Analysis Antibacterial agents Antitoxins Bacteria Bacterial Proteins - genetics Bacteriocins Base sequence Base Sequence - genetics Biological products Biology and life sciences Chloramphenicol Chloromycetin Chromosomes Cloning Copy number Deoxyribonucleic acid DNA DNA Replication - genetics DNA, Bacterial - genetics E coli Electroporation Gene expression Genes Genetic engineering Genetic vectors Genetic Vectors - genetics Genomes Host range Identity Integrase Lactobacillus Lactobacillus - genetics Lactobacillus curvatus Life Sciences Medicine and Health Sciences Nucleotide sequence Nucleotides Phenols (Class of compounds) Plasmids Plasmids - genetics Recombinants RepA gene Replicating Replication Replication Origin - genetics Replication origins Replicon - genetics Research and analysis methods Sequence Analysis, DNA - methods Toxins Transposases - genetics Tyrosine Vectors (Biology) |
| title | Nucleotide sequence and analysis of pRC12 and pRC18, two theta-replicating plasmids harbored by Lactobacillus curvatus CRL 705 |
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