Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro

Detalhes bibliográficos
Autor(a) principal: Bastos, Matheus Silva e
Data de Publicação: 2016
Outros Autores: Souza, Luciana Ângelo de, Onofre, Thiago Souza, Almeida, Márcia Rogéria de, Bressan, Gustavo Costa, Fietto, Juliana Lopes Rangel, Silva Júnior, Abelardo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://dx.doi.org/10.1590/0074-02760160237
http://www.locus.ufv.br/handle/123456789/12018
Resumo: Gene reporter-fluorescent cells have emerged as alternative method for drug screening. Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.
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spelling Bastos, Matheus Silva eSouza, Luciana Ângelo deOnofre, Thiago SouzaAlmeida, Márcia Rogéria deBressan, Gustavo CostaFietto, Juliana Lopes RangelSilva Júnior, Abelardo2017-10-11T13:47:46Z2017-10-11T13:47:46Z2016-10-201678-8060http://dx.doi.org/10.1590/0074-02760160237http://www.locus.ufv.br/handle/123456789/12018Gene reporter-fluorescent cells have emerged as alternative method for drug screening. Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.engMemórias do Instituto Oswaldo Cruzvol.112 n.2, 155-159 Feb. 2017Leishmania braziliensisGreen fluorescent proteinDrug screeningAchievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitroinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINAL0074-0276-mioc-112-2-0155.pdf0074-0276-mioc-112-2-0155.pdftexto completoapplication/pdf790870https://locus.ufv.br//bitstream/123456789/12018/1/0074-0276-mioc-112-2-0155.pdfd3cc400aeddf56b3e392e09901676e6eMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://locus.ufv.br//bitstream/123456789/12018/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52THUMBNAIL0074-0276-mioc-112-2-0155.pdf.jpg0074-0276-mioc-112-2-0155.pdf.jpgIM Thumbnailimage/jpeg7212https://locus.ufv.br//bitstream/123456789/12018/3/0074-0276-mioc-112-2-0155.pdf.jpg598b90906fee874a8b96a9301dcf9925MD53123456789/120182017-10-11 23:00:48.297oai:locus.ufv.br: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Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452017-10-12T02:00:48LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.en.fl_str_mv Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
title Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
spellingShingle Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
Bastos, Matheus Silva e
Leishmania braziliensis
Green fluorescent protein
Drug screening
title_short Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
title_full Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
title_fullStr Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
title_full_unstemmed Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
title_sort Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro
author Bastos, Matheus Silva e
author_facet Bastos, Matheus Silva e
Souza, Luciana Ângelo de
Onofre, Thiago Souza
Almeida, Márcia Rogéria de
Bressan, Gustavo Costa
Fietto, Juliana Lopes Rangel
Silva Júnior, Abelardo
author_role author
author2 Souza, Luciana Ângelo de
Onofre, Thiago Souza
Almeida, Márcia Rogéria de
Bressan, Gustavo Costa
Fietto, Juliana Lopes Rangel
Silva Júnior, Abelardo
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Bastos, Matheus Silva e
Souza, Luciana Ângelo de
Onofre, Thiago Souza
Almeida, Márcia Rogéria de
Bressan, Gustavo Costa
Fietto, Juliana Lopes Rangel
Silva Júnior, Abelardo
dc.subject.pt-BR.fl_str_mv Leishmania braziliensis
Green fluorescent protein
Drug screening
topic Leishmania braziliensis
Green fluorescent protein
Drug screening
description Gene reporter-fluorescent cells have emerged as alternative method for drug screening. Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.
publishDate 2016
dc.date.issued.fl_str_mv 2016-10-20
dc.date.accessioned.fl_str_mv 2017-10-11T13:47:46Z
dc.date.available.fl_str_mv 2017-10-11T13:47:46Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1590/0074-02760160237
http://www.locus.ufv.br/handle/123456789/12018
dc.identifier.issn.none.fl_str_mv 1678-8060
identifier_str_mv 1678-8060
url http://dx.doi.org/10.1590/0074-02760160237
http://www.locus.ufv.br/handle/123456789/12018
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartofseries.pt-BR.fl_str_mv vol.112 n.2, 155-159 Feb. 2017
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Memórias do Instituto Oswaldo Cruz
publisher.none.fl_str_mv Memórias do Instituto Oswaldo Cruz
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
instname:Universidade Federal de Viçosa (UFV)
instacron:UFV
instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
reponame_str LOCUS Repositório Institucional da UFV
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