Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi

Detalhes bibliográficos
Autor(a) principal: Nora, Luísa Czamanski
Data de Publicação: 2019
Outros Autores: Gonçales, Relber Aguiar, Martins-Santana, Leonardo, Ferreira, Beatriz Henriques, Rodrigues, Fernando José dos Santos, Silva-Rocha, Rafael
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/62304
Resumo: We present a collection of minimalist binary vectors for transformation through ATMT applicable to several fungi species. pLUO plasmid binary vectors consist of a reporter module containing fluorescent proteins, mCherry or eGFP, flanked by a multiple cloning site and a transcription terminator site. They also present a synthetic gene allowing resistance to Hygromicin B flanked by alternate promoters, one for yeast and another for filamentous fungi. Left and right borders were added for Agrobacterium tumefaciens recognition, and a minimal broad-host range RK2 replication origin. Transformation was validated in the pathogenic fungus Paracoccidioides lutzii. Hence, we developed an efficient and reliable molecular tool for fungal transformation: minimalist, synthetic, modular, and available in four different versions, and these can still be readily modified using a few primers and few cloning steps.
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spelling Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungiSynthetic biologyFungiAgrobacterium tumefaciensTransformationVectorsScience & TechnologyWe present a collection of minimalist binary vectors for transformation through ATMT applicable to several fungi species. pLUO plasmid binary vectors consist of a reporter module containing fluorescent proteins, mCherry or eGFP, flanked by a multiple cloning site and a transcription terminator site. They also present a synthetic gene allowing resistance to Hygromicin B flanked by alternate promoters, one for yeast and another for filamentous fungi. Left and right borders were added for Agrobacterium tumefaciens recognition, and a minimal broad-host range RK2 replication origin. Transformation was validated in the pathogenic fungus Paracoccidioides lutzii. Hence, we developed an efficient and reliable molecular tool for fungal transformation: minimalist, synthetic, modular, and available in four different versions, and these can still be readily modified using a few primers and few cloning steps.LCN, LMS, RAG and RSR were supported by FAPESP (Project numbers: 2016/03763-3, 2016/01946-3, 2014/22561-7 and 2012/22921-8). FR and BF were supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) (NORTE-01-0145-FEDER000013).Sociedade Brasileira de GenéticaUniversidade do MinhoNora, Luísa CzamanskiGonçales, Relber AguiarMartins-Santana, LeonardoFerreira, Beatriz HenriquesRodrigues, Fernando José dos SantosSilva-Rocha, Rafael20192019-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/62304eng1415-47571678-468510.1590/1678-4685-GMB-2018-0221info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:07:36ZPortal AgregadorONG
dc.title.none.fl_str_mv Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
title Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
spellingShingle Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
Nora, Luísa Czamanski
Synthetic biology
Fungi
Agrobacterium tumefaciens
Transformation
Vectors
Science & Technology
title_short Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
title_full Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
title_fullStr Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
title_full_unstemmed Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
title_sort Synthetic and minimalist vectors for Agrobacterium tumefaciens-mediated transformation of fungi
author Nora, Luísa Czamanski
author_facet Nora, Luísa Czamanski
Gonçales, Relber Aguiar
Martins-Santana, Leonardo
Ferreira, Beatriz Henriques
Rodrigues, Fernando José dos Santos
Silva-Rocha, Rafael
author_role author
author2 Gonçales, Relber Aguiar
Martins-Santana, Leonardo
Ferreira, Beatriz Henriques
Rodrigues, Fernando José dos Santos
Silva-Rocha, Rafael
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Nora, Luísa Czamanski
Gonçales, Relber Aguiar
Martins-Santana, Leonardo
Ferreira, Beatriz Henriques
Rodrigues, Fernando José dos Santos
Silva-Rocha, Rafael
dc.subject.por.fl_str_mv Synthetic biology
Fungi
Agrobacterium tumefaciens
Transformation
Vectors
Science & Technology
topic Synthetic biology
Fungi
Agrobacterium tumefaciens
Transformation
Vectors
Science & Technology
description We present a collection of minimalist binary vectors for transformation through ATMT applicable to several fungi species. pLUO plasmid binary vectors consist of a reporter module containing fluorescent proteins, mCherry or eGFP, flanked by a multiple cloning site and a transcription terminator site. They also present a synthetic gene allowing resistance to Hygromicin B flanked by alternate promoters, one for yeast and another for filamentous fungi. Left and right borders were added for Agrobacterium tumefaciens recognition, and a minimal broad-host range RK2 replication origin. Transformation was validated in the pathogenic fungus Paracoccidioides lutzii. Hence, we developed an efficient and reliable molecular tool for fungal transformation: minimalist, synthetic, modular, and available in four different versions, and these can still be readily modified using a few primers and few cloning steps.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-01-01T00:00:00Z
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://hdl.handle.net/1822/62304
url http://hdl.handle.net/1822/62304
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1415-4757
1678-4685
10.1590/1678-4685-GMB-2018-0221
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 Sociedade Brasileira de Genética
publisher.none.fl_str_mv Sociedade Brasileira de Genética
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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