pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation

Detalhes bibliográficos
Autor(a) principal: Kugeratski,Fernanda G
Data de Publicação: 2015
Outros Autores: Batista,Michel, Inoue,Alexandre Haruo, Ramos,Bruno Dias, Krieger,Marco Aurelio, Marchini/,Fabricio K
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Memórias do Instituto Oswaldo Cruz
Texto Completo: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000500687
Resumo: The functional characterisation of thousands of Trypanosoma cruzi genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the pTcGW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway® technology. Here, we present an improved 1.1 version of pTcGW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four T. cruzi proteins. The 1.1 version of pTcGW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to T. cruzi genes and is freely available for scientific community.
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spelling pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisationTrypanosoma cruziplasmid vectorspTcGW platformreverse geneticscloninggene characterisationThe functional characterisation of thousands of Trypanosoma cruzi genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the pTcGW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway® technology. Here, we present an improved 1.1 version of pTcGW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four T. cruzi proteins. The 1.1 version of pTcGW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to T. cruzi genes and is freely available for scientific community.Instituto Oswaldo Cruz, Ministério da Saúde2015-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000500687Memórias do Instituto Oswaldo Cruz v.110 n.5 2015reponame:Memórias do Instituto Oswaldo Cruzinstname:Fundação Oswaldo Cruzinstacron:FIOCRUZ10.1590/0074-02760150074info:eu-repo/semantics/openAccessKugeratski,Fernanda GBatista,MichelInoue,Alexandre HaruoRamos,Bruno DiasKrieger,Marco AurelioMarchini/,Fabricio Keng2020-04-25T17:52:17Zhttp://www.scielo.br/oai/scielo-oai.php0074-02761678-8060opendoar:null2020-04-26 02:20:49.937Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruztrue
dc.title.none.fl_str_mv pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
title pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
spellingShingle pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
Kugeratski,Fernanda G
Trypanosoma cruzi
plasmid vectors
pTcGW platform
reverse genetics
cloning
gene characterisation
title_short pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
title_full pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
title_fullStr pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
title_full_unstemmed pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
title_sort pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma cruzi gene characterisation
author Kugeratski,Fernanda G
author_facet Kugeratski,Fernanda G
Batista,Michel
Inoue,Alexandre Haruo
Ramos,Bruno Dias
Krieger,Marco Aurelio
Marchini/,Fabricio K
author_role author
author2 Batista,Michel
Inoue,Alexandre Haruo
Ramos,Bruno Dias
Krieger,Marco Aurelio
Marchini/,Fabricio K
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Kugeratski,Fernanda G
Batista,Michel
Inoue,Alexandre Haruo
Ramos,Bruno Dias
Krieger,Marco Aurelio
Marchini/,Fabricio K
dc.subject.por.fl_str_mv Trypanosoma cruzi
plasmid vectors
pTcGW platform
reverse genetics
cloning
gene characterisation
topic Trypanosoma cruzi
plasmid vectors
pTcGW platform
reverse genetics
cloning
gene characterisation
dc.description.none.fl_txt_mv The functional characterisation of thousands of Trypanosoma cruzi genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the pTcGW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway® technology. Here, we present an improved 1.1 version of pTcGW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four T. cruzi proteins. The 1.1 version of pTcGW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to T. cruzi genes and is freely available for scientific community.
description The functional characterisation of thousands of Trypanosoma cruzi genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the pTcGW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway® technology. Here, we present an improved 1.1 version of pTcGW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four T. cruzi proteins. The 1.1 version of pTcGW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to T. cruzi genes and is freely available for scientific community.
publishDate 2015
dc.date.none.fl_str_mv 2015-08-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000500687
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000500687
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/0074-02760150074
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Instituto Oswaldo Cruz, Ministério da Saúde
publisher.none.fl_str_mv Instituto Oswaldo Cruz, Ministério da Saúde
dc.source.none.fl_str_mv Memórias do Instituto Oswaldo Cruz v.110 n.5 2015
reponame:Memórias do Instituto Oswaldo Cruz
instname:Fundação Oswaldo Cruz
instacron:FIOCRUZ
reponame_str Memórias do Instituto Oswaldo Cruz
collection Memórias do Instituto Oswaldo Cruz
instname_str Fundação Oswaldo Cruz
instacron_str FIOCRUZ
institution FIOCRUZ
repository.name.fl_str_mv Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruz
repository.mail.fl_str_mv
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