Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tde/1297 |
Resumo: | Samonella spp are Gram negatives bactérias belonging to Enterobacteriaceae family. S. enterica comprise about 2.500 sorovars. These sorovars can infect a broad range, including poultry, cattle, swins and humans, and are agent causative of salmonellosis an important public health issue worldwide. Small multicopy plasmids are frequently isolated from Gram negatives and Gram positives bacterias. In Salmonella, low molecular weight plasmids are found on 10% of Salmonella strains and their biological functions are unknown. However, many plasmids in Salmonella control important properties, such as, virulence factors, heavy metals and antibiotics resistance, and utilizations of alternative carbon sources. The pVCM1 plasmid was extracted from one strain of Samonella enteretidis isolated from broilers carcass. The strains were grown in liquid or solid Luria-Bertani broth at 37 °C. The plasmid was purified, separated on 1% agarose electrophoresis and visualized by ethidium bromide staining for analysis. Plasmid was digested with EcoRI enzyme and subcloned in the pUC18 vector.The plasmidal stability was evaluated, inoculating E. coli cells transformated with pVCM1 plasmid (cloned in pUC18) in liquid Luria-bertani broth supplemented with ampicillin. The pVCM1 was stable after 240 generations. The total DNA sequence of plasmid pVCM1 has 1981 pb. Genbank search resulted that pVCM1 showed 99% of identity with pB and 92% with pJ, which were isolated from Salmonella enteretidis. Only one ORF founded in pVCM1 showed significative similarity with others proteins of GenBank. The protein encoded by this ORF showed homology to Rep proteins of plasmids that replicates by rolling-circle mechanism. The pVCM1 posses three impotents elements: rep gene, single strand origin (SSO) and inverted repeat sequences. Such elements are importants for the rolling circle replication, suggesting that pVCM1 use this mechanism. The rep gene was amplified and cloned in the pGEMT-easy vector, but the heterologous expression of Rep protein wasn t gotten successfully. |
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BATAUS, Luiz Artur Mendeshttp://lattes.cnpq.br/5637230378599476http://lattes.cnpq.br/0324512679542455PENIDO, Ana Flávia Batista2014-07-29T15:16:38Z2010-08-232009-03-26PENIDO, Ana Flávia Batista. Complete sequence and carachterization of cryptic plasmid isoladed from Salmonella enterica. 2009. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2009.http://repositorio.bc.ufg.br/tede/handle/tde/1297ark:/38995/001300000d6f7Samonella spp are Gram negatives bactérias belonging to Enterobacteriaceae family. S. enterica comprise about 2.500 sorovars. These sorovars can infect a broad range, including poultry, cattle, swins and humans, and are agent causative of salmonellosis an important public health issue worldwide. Small multicopy plasmids are frequently isolated from Gram negatives and Gram positives bacterias. In Salmonella, low molecular weight plasmids are found on 10% of Salmonella strains and their biological functions are unknown. However, many plasmids in Salmonella control important properties, such as, virulence factors, heavy metals and antibiotics resistance, and utilizations of alternative carbon sources. The pVCM1 plasmid was extracted from one strain of Samonella enteretidis isolated from broilers carcass. The strains were grown in liquid or solid Luria-Bertani broth at 37 °C. The plasmid was purified, separated on 1% agarose electrophoresis and visualized by ethidium bromide staining for analysis. Plasmid was digested with EcoRI enzyme and subcloned in the pUC18 vector.The plasmidal stability was evaluated, inoculating E. coli cells transformated with pVCM1 plasmid (cloned in pUC18) in liquid Luria-bertani broth supplemented with ampicillin. The pVCM1 was stable after 240 generations. The total DNA sequence of plasmid pVCM1 has 1981 pb. Genbank search resulted that pVCM1 showed 99% of identity with pB and 92% with pJ, which were isolated from Salmonella enteretidis. Only one ORF founded in pVCM1 showed significative similarity with others proteins of GenBank. The protein encoded by this ORF showed homology to Rep proteins of plasmids that replicates by rolling-circle mechanism. The pVCM1 posses three impotents elements: rep gene, single strand origin (SSO) and inverted repeat sequences. Such elements are importants for the rolling circle replication, suggesting that pVCM1 use this mechanism. The rep gene was amplified and cloned in the pGEMT-easy vector, but the heterologous expression of Rep protein wasn t gotten successfully.Salmonellas ssp são bactérias Gram negativas pertencente a família das enterobácterias. A S. entérica compreende cerca de 2.500 sorovares. Esses sorovares podem infectar vários hospedeiros incluindo aves, bovinos, suínos e humanos, e são os agentes causais das salmoneloses, um importante problema de saúde pública em todo mundo. Múltiplas cópias de pequenos plasmídeos são frequentemente isolados de bactérias Gram negativas e Gram positivas. Nas salmonelas, plasmídeos com baixa massa molecular são encontradas em 10% das linhagens e suas funções biológicas são desconhecidas. Entretanto, muitos plasmídeos de salmonela controlam propriedades importantes, tais como, fatores de virulência, resistência a antibióticos e metais pesados, e utilização de fontes alternativas de carbono. O plasmídeo pVCM1 foi extraído de uma linhagem S. Enteritidis, isoladas de carcaças de frango. Essa linhagem foi crescida em meio Lúria-Bertani líquido e sólido à 37°C. O plasmídeo foi purificado, separado em gel de agarose 1% e visualizado com coloração por brometo de etídio. O plasmídeo foi digerido com EcoRI e subclonado no vetor pUC18. A estabilidade plasmidial foi avaliada inoculando células de E. coli transformadas com plasmídeo pVCM1 (clonado em pUC18) em meio Lúria- bertani líquido suplementado com ampicilina. O pVCM1 foi estável por mais de 240 gerações A sequência nucleotídica total do plasmídeo possui 1981 pb. Pequisa por sequências no GenBank, mostrou que o pVCM1 apresenta similaridade de 99% com o plasmídeo pB e 92% com plasmídeo pJ, os quais foram isolados de S. Enteritidis.Das 11 possíveis ORFS apenas uma única ORF apresentou similaridade significativa com outras proteínas do GenBank. A proteína codificada por essa ORF apresentou homologia com as proteínas Rep de plasmídeos que replicam via círculo rolante. O pVCM1 apresenta três regiões importantes: gene rep, origem de fita simples (SSO) e regiões repetidas invertidas. Tais regiões são importantes para o mecanismo de replicação via círculo rolante, sugerindo que o pVCM1 utilize esse mecanismo. O gene rep foi amplificado e clonado no vetor pGEM T-easy, mas a expressão heteróloga da proteína Rep não foi obtida com sucesso.Made available in DSpace on 2014-07-29T15:16:38Z (GMT). No. of bitstreams: 1 Dissertacao Ana Flavia B Penido.pdf: 829056 bytes, checksum: 63d111404f5b1f9b2d136e54aad128a4 (MD5) Previous issue date: 2009-03-26application/pdfhttp://repositorio.bc.ufg.br/TEDE/retrieve/3975/Dissertacao%20Ana%20Flavia%20B%20%20Penido.pdf.jpgporUniversidade Federal de GoiásMestrado em BiologiaUFGBRCiências Biolóicas1. Plasmídeo; 2. Salmonella entérica 3. Replicação via círculo-rolante1. Plasmídeo2. Replicação via círculo-rolante3. Salmonella1. Plasmid2. Rolling-circle replication3. SalmonellaCNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERALSequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella entericaComplete sequence and carachterization of cryptic plasmid isoladed from Salmonella entericainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGORIGINALDissertacao Ana Flavia B Penido.pdfapplication/pdf829056http://repositorio.bc.ufg.br/tede/bitstreams/2244f7bc-585d-4dbe-bcdd-2825a9a167bc/download63d111404f5b1f9b2d136e54aad128a4MD51TEXTDissertacao Ana Flavia B Penido.pdf.txtDissertacao Ana Flavia B Penido.pdf.txtExtracted Texttext/plain155445http://repositorio.bc.ufg.br/tede/bitstreams/20b3c9d2-90ac-4b19-89e3-e3aa39c4c9d5/download81a1775830827248addd3eb5146ea6b0MD52THUMBNAILDissertacao Ana Flavia B Penido.pdf.jpgDissertacao Ana Flavia B Penido.pdf.jpgGenerated Thumbnailimage/jpeg2965http://repositorio.bc.ufg.br/tede/bitstreams/49517607-4a1e-4595-b6c0-af500785f363/downloadfed04d7dc5d6880e3edd320432be8e61MD53tde/12972014-07-30 03:09:50.462open.accessoai:repositorio.bc.ufg.br:tde/1297http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2014-07-30T06:09:50Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)false |
dc.title.por.fl_str_mv |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
dc.title.alternative.eng.fl_str_mv |
Complete sequence and carachterization of cryptic plasmid isoladed from Salmonella enterica |
title |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
spellingShingle |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica PENIDO, Ana Flávia Batista 1. Plasmídeo; 2. Salmonella entérica 3. Replicação via círculo-rolante 1. Plasmídeo 2. Replicação via círculo-rolante 3. Salmonella 1. Plasmid 2. Rolling-circle replication 3. Salmonella CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
title_full |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
title_fullStr |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
title_full_unstemmed |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
title_sort |
Sequência completa e caracterização do plasmídeo crípico pVCM1 isolado de Salmonella enterica |
author |
PENIDO, Ana Flávia Batista |
author_facet |
PENIDO, Ana Flávia Batista |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
BATAUS, Luiz Artur Mendes |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5637230378599476 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/0324512679542455 |
dc.contributor.author.fl_str_mv |
PENIDO, Ana Flávia Batista |
contributor_str_mv |
BATAUS, Luiz Artur Mendes |
dc.subject.por.fl_str_mv |
1. Plasmídeo; 2. Salmonella entérica 3. Replicação via círculo-rolante 1. Plasmídeo 2. Replicação via círculo-rolante 3. Salmonella |
topic |
1. Plasmídeo; 2. Salmonella entérica 3. Replicação via círculo-rolante 1. Plasmídeo 2. Replicação via círculo-rolante 3. Salmonella 1. Plasmid 2. Rolling-circle replication 3. Salmonella CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
dc.subject.eng.fl_str_mv |
1. Plasmid 2. Rolling-circle replication 3. Salmonella |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
Samonella spp are Gram negatives bactérias belonging to Enterobacteriaceae family. S. enterica comprise about 2.500 sorovars. These sorovars can infect a broad range, including poultry, cattle, swins and humans, and are agent causative of salmonellosis an important public health issue worldwide. Small multicopy plasmids are frequently isolated from Gram negatives and Gram positives bacterias. In Salmonella, low molecular weight plasmids are found on 10% of Salmonella strains and their biological functions are unknown. However, many plasmids in Salmonella control important properties, such as, virulence factors, heavy metals and antibiotics resistance, and utilizations of alternative carbon sources. The pVCM1 plasmid was extracted from one strain of Samonella enteretidis isolated from broilers carcass. The strains were grown in liquid or solid Luria-Bertani broth at 37 °C. The plasmid was purified, separated on 1% agarose electrophoresis and visualized by ethidium bromide staining for analysis. Plasmid was digested with EcoRI enzyme and subcloned in the pUC18 vector.The plasmidal stability was evaluated, inoculating E. coli cells transformated with pVCM1 plasmid (cloned in pUC18) in liquid Luria-bertani broth supplemented with ampicillin. The pVCM1 was stable after 240 generations. The total DNA sequence of plasmid pVCM1 has 1981 pb. Genbank search resulted that pVCM1 showed 99% of identity with pB and 92% with pJ, which were isolated from Salmonella enteretidis. Only one ORF founded in pVCM1 showed significative similarity with others proteins of GenBank. The protein encoded by this ORF showed homology to Rep proteins of plasmids that replicates by rolling-circle mechanism. The pVCM1 posses three impotents elements: rep gene, single strand origin (SSO) and inverted repeat sequences. Such elements are importants for the rolling circle replication, suggesting that pVCM1 use this mechanism. The rep gene was amplified and cloned in the pGEMT-easy vector, but the heterologous expression of Rep protein wasn t gotten successfully. |
publishDate |
2009 |
dc.date.issued.fl_str_mv |
2009-03-26 |
dc.date.available.fl_str_mv |
2010-08-23 |
dc.date.accessioned.fl_str_mv |
2014-07-29T15:16:38Z |
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info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
PENIDO, Ana Flávia Batista. Complete sequence and carachterization of cryptic plasmid isoladed from Salmonella enterica. 2009. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2009. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tde/1297 |
dc.identifier.dark.fl_str_mv |
ark:/38995/001300000d6f7 |
identifier_str_mv |
PENIDO, Ana Flávia Batista. Complete sequence and carachterization of cryptic plasmid isoladed from Salmonella enterica. 2009. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2009. ark:/38995/001300000d6f7 |
url |
http://repositorio.bc.ufg.br/tede/handle/tde/1297 |
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por |
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openAccess |
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Universidade Federal de Goiás |
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UFG |
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BR |
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Ciências Biolóicas |
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Universidade Federal de Goiás |
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