A new vector for heterologous gene expression in Escherichia coli with increased stability in the absence of antibiotic

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dc.contributorLaboratório Especial de Desenvolvimento de Vacinas (LEDV)pt_BR
dc.contributorLaboratório de Bacteriologiapt_BR
dc.contributorCentro Bioindustrialpt_BR
dc.contributor.authorEguia, Fara Amelia Primellespt_BR
dc.contributor.authorRamos, Henrique Romanpt_BR
dc.contributor.authorKraschowetz, Stefaniept_BR
dc.contributor.authorOmote, Daniel de Queirozpt_BR
dc.contributor.authorRamos, Celso Raúl Romeropt_BR
dc.contributor.authorHo, Paulo Leept_BR
dc.contributor.authorCarvalho, Eneaspt_BR
dc.contributor.authorGonçalves, Viviane Maimonipt_BR
dc.date.accessioned2020-07-09T21:21:25Z-
dc.date.available2020-07-09T21:21:25Z-
dc.date.issued2018-
dc.identifier.citationEguia FAP, Ramos HR, Kraschowetz S, Omote DQ, Ramos CRR, Ho PL, et al. A new vector for heterologous gene expression in Escherichia coli with increased stability in the absence of antibiotic. Plasmid. 2018 Jun;98:22-30. doi:10.1016/j.plasmid.2018.08.004.pt_BR
dc.identifier.issn0147-619X-
dc.identifier.issn1095-9890-
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2569-
dc.description.abstractExpression vectors for industrial production should be stable and allow tight control of protein synthesis. This is necessary to ensure plasmid transmission to daughter cells in order to achieve a stable population capable of synthesizing high amounts of the target protein. A high-copy-number plasmid, pAE, was previously used for laboratory-scale production of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and the Schistosoma mansoni fatty acid binding protein (rSm14), but it was unstable for large-scale production. Therefore, here we evaluated a new expression vector derived from pAE, pAR-KanI, which combines two plasmid replication strategies: a high-copy plasmid pUC origin of replication as pAE, and a par locus sequence derived from pSC101, which is typical of low copy plasmids, for rhG-CSF and rSm14 production in Escherichia coli. Clones bearing these constructs were cultivated in two complex media (2YT and auto-induction) and both yielded higher-than-95% resistant colonies, before and after induction, either with or without antibiotics. In 2YT medium, we obtained 244?µg/mL of rSm14, 181?µg/mL and 392?µg/mL for rhG-CSF, with and without glucose, respectively. In auto-induction medium without antibiotics, 147?µg/mL of rSm14 and 162?µg/mL of rhG-CSF were obtained. The new vector presented high stability for the production of both recombinant proteins in complex media in Escherichia coli, even in the absence of antibiotics, making the pAR-KanI a promising vector for industrial production of recombinant proteins.pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)pt_BR
dc.format.extentp. 22-30pt_BR
dc.languageengpt_BR
dc.publisherAcademic Presspt_BR
dc.relation.ispartofPlasmidpt_BR
dc.titleA new vector for heterologous gene expression in Escherichia coli with increased stability in the absence of antibioticpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1016/j.plasmid.2018.08.004pt_BR
dc.identifier.urlhttps://doi.org/10.1016/j.plasmid.2018.08.004pt_BR
dc.contributor.externalUniversidade Nove de Julho (UNINOVE)¦¦Brasilpt_BR
dc.publisher.cityNew Yorkpt_BR
dc.identifier.citationvolume98pt_BR
dc.subject.keywordrhG-CSFpt_BR
dc.subject.keywordNartograstimpt_BR
dc.subject.keywordSm14pt_BR
dc.subject.keywordRecombinant Escherichia colipt_BR
dc.subject.keywordPlasmid stabilitypt_BR
dc.subject.keywordAntibiotic resistancept_BR
dc.relation.ispartofabbreviatedPlasmidpt_BR
dc.identifier.citationabntv. 98, p. 22-30, jun. 2018pt_BR
dc.identifier.citationvancouver2018 Jun;98:22-30pt_BR
dc.publisher.countryUnited Statespt_BR
dc.contributor.butantanRamos, Henrique Roman|:Aluno|:Laboratório de Bacteriologia|:pt_BR
dc.contributor.butantanKraschowetz, Stefanie|:Aluno|:Laboratório Especial de Desenvolvimento de Vacinas (LEDV)|:pt_BR
dc.contributor.butantanOmote, Daniel de Queiroz|:Aluno|:Laboratório de Bacteriologia|:pt_BR
dc.contributor.butantanRamos, Celso Raúl Romero|:Aluno|:Centro de Biotecnologia|:pt_BR
dc.contributor.butantanCarvalho, Eneas|:Pesquisador|:Laboratório de Bacteriologia|:pt_BR
dc.contributor.butantanGonçalves, Viviane Maimoni|:Pesquisador|:Laboratório Especial de Desenvolvimento de Vacinas (LEDV)|:Autor de correspondênciapt_BR
dc.contributor.butantanHo, Paulo Lee|:Pesquisador|:Centro Bioindustrial|:pt_BR
dc.contributor.butantanEguia, Fara Amelia Primelles|:Aluno|:Laboratório Especial de Desenvolvimento de Vacinas (LEDV)|:PrimeiroAutorpt_BR
dc.sponsorship.butantanConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)¦¦118669/2011-7pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦12585-13-0pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
item.openairetypeArticle-
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item.grantfulltextembargo_29990101-
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