Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae

Detalhes bibliográficos
Autor(a) principal: Pereira, Filipa
Data de Publicação: 2010
Outros Autores: Vieira, N., Casal, Margarida, Alistair, Brown, Johansson, Björn, Paiva, Sandra
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/16014
Resumo: A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli possibly because of their toxic effects. The ligation of S. cerevisiae 2μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors CIp10, CIp20 and CIp30 are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nº: CIp10-2μ (GU550119), CIp20-2μ (GU550120) and CIp30-2μ (GU550121).
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spelling Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiaeCandida albicansPlasmidsyeastgap-repairtoxic genesplasmid integrationin vivoScience & TechnologyA general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli possibly because of their toxic effects. The ligation of S. cerevisiae 2μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors CIp10, CIp20 and CIp30 are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nº: CIp10-2μ (GU550119), CIp20-2μ (GU550120) and CIp30-2μ (GU550121).This study was supported by FEDER, Portugal (Grant No. POCI/BIA-BCM/57812/2004; Eixo 2, Medida 45 2.3, QCAIII-FEDER). N.V. received a FCT PhD fellowship (Grant No. SFRH/BD/23503/2005). F. P. and B.J. were supported by European Union project NILE (Grant No. FP6-019882). AJPB was funded by the BBSRC (Grant Nos BB/F000111/1, BB/D009308/1, BB/F010826/1 and BB/FO0513X/1) and the European Commission (Grant Nos PITN-GA-2008-214004 and ERC-2009-AdG-249793).Wiley-BlackwellUniversidade do MinhoPereira, FilipaVieira, N.Casal, MargaridaAlistair, BrownJohansson, BjörnPaiva, Sandra20102010-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/16014eng0749-503X10.1002/yea.180020602447info: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:RCAAP2023-07-21T12:51:22Zoai:repositorium.sdum.uminho.pt:1822/16014Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:50:15.714717Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
title Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
spellingShingle Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
Pereira, Filipa
Candida albicans
Plasmids
yeast
gap-repair
toxic genes
plasmid integration
in vivo
Science & Technology
title_short Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
title_full Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
title_fullStr Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
title_full_unstemmed Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
title_sort Plasmids for in vivo construction of integrative Candida albicans vectors in saccharomyces cerevisiae
author Pereira, Filipa
author_facet Pereira, Filipa
Vieira, N.
Casal, Margarida
Alistair, Brown
Johansson, Björn
Paiva, Sandra
author_role author
author2 Vieira, N.
Casal, Margarida
Alistair, Brown
Johansson, Björn
Paiva, Sandra
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Pereira, Filipa
Vieira, N.
Casal, Margarida
Alistair, Brown
Johansson, Björn
Paiva, Sandra
dc.subject.por.fl_str_mv Candida albicans
Plasmids
yeast
gap-repair
toxic genes
plasmid integration
in vivo
Science & Technology
topic Candida albicans
Plasmids
yeast
gap-repair
toxic genes
plasmid integration
in vivo
Science & Technology
description A general system has been devised for the in vivo construction of Candida albicans integrative vectors in Saccharomyces cerevisiae. The system is especially useful for the integration of genes in C. albicans that cannot be propagated in Escherichia coli possibly because of their toxic effects. The ligation of S. cerevisiae 2μ sequences to a C. albicans integrative vector permits in vivo maintenance and gap repair cloning within S. cerevisiae. After the vector assembly, it can be purified from S. cerevisiae or amplified by PCR and then used for transformation of C. albicans. The S. cerevisiae 2μ sequence is completely removed by linearization prior to C. albicans transformation, such that no unwanted DNA is transferred in the final construct. The system was successfully used to clone and reintegrate the C. albicans JEN2 gene, which encodes a membrane protein that is apparently toxic to E. coli. Three popular C. albicans integrative vectors CIp10, CIp20 and CIp30 are now available in versions that permit gap repair in S. cerevisiae. GenBank Accession Nº: CIp10-2μ (GU550119), CIp20-2μ (GU550120) and CIp30-2μ (GU550121).
publishDate 2010
dc.date.none.fl_str_mv 2010
2010-01-01T00:00:00Z
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/1822/16014
url http://hdl.handle.net/1822/16014
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0749-503X
10.1002/yea.1800
20602447
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 Wiley-Blackwell
publisher.none.fl_str_mv Wiley-Blackwell
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
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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