Cloning of two new CDPK isoforms from Solanum tuberosum

Autores
Fantino, Elisa Inés; Santin, Franco; Ulloa, Rita Maria
Año de publicación
2013
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Calcium-dependent protein kinases (CDPKs) comprise a multigene family of calcium sensors that play important roles regulating plant growth and development and plant responses to environmental stresses. To the moment six isoforms have been characterized in potato plants. In silico analysis of Solanum phureja genome allowed us to identify other 20 candidate genes encoding potential CDPK isoforms that cluster into four subgroups. Expression of 13 of these isoforms was detected in leaves and stolons from Solanum tuberosum cv Desirée plants. Using primers directed against their 5´and 3´utr sequences we amplified isoforms StCDPK25 and StCDPK17 belonging to subgroup 1 and 3 respectively. These fragments were cloned and sequenced. Isoform StCDPK25 shares 99% identity with the S. phureja coding sequence and presented all the characteristics of a CDPK. Cloning of StCDPK25 coding sequence in pDEST expression vector in order to obtain the recombinant enzyme tagged to 6xHis is currently in progress. Analysis of its sequence indicated that it could be targeted to chloroplasts; the N-terminal sequence will be fused to GFP and transient expression assays will be performed in Nicotiana benthamiana. On the contrary, only a 1 kb fragment containing exons 1, 2 and 7 was amplified for StCDPK17 suggesting that alternative splicing has occurred.
Fil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Santin, Franco. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Molecular mechanisms in cell signaling and gene expression
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
Materia
CDPKs
CALCIO
SOLANUM TUBEROSUM
CHLOROPLASTS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/244644
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/244644
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cloning of two new CDPK isoforms from Solanum tuberosumFantino, Elisa InésSantin, FrancoUlloa, Rita MariaCDPKsCALCIOSOLANUM TUBEROSUMCHLOROPLASTShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Calcium-dependent protein kinases (CDPKs) comprise a multigene family of calcium sensors that play important roles regulating plant growth and development and plant responses to environmental stresses. To the moment six isoforms have been characterized in potato plants. In silico analysis of Solanum phureja genome allowed us to identify other 20 candidate genes encoding potential CDPK isoforms that cluster into four subgroups. Expression of 13 of these isoforms was detected in leaves and stolons from Solanum tuberosum cv Desirée plants. Using primers directed against their 5´and 3´utr sequences we amplified isoforms StCDPK25 and StCDPK17 belonging to subgroup 1 and 3 respectively. These fragments were cloned and sequenced. Isoform StCDPK25 shares 99% identity with the S. phureja coding sequence and presented all the characteristics of a CDPK. Cloning of StCDPK25 coding sequence in pDEST expression vector in order to obtain the recombinant enzyme tagged to 6xHis is currently in progress. Analysis of its sequence indicated that it could be targeted to chloroplasts; the N-terminal sequence will be fused to GFP and transient expression assays will be performed in Nicotiana benthamiana. On the contrary, only a 1 kb fragment containing exons 1, 2 and 7 was amplified for StCDPK17 suggesting that alternative splicing has occurred.Fil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Santin, Franco. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaMolecular mechanisms in cell signaling and gene expressionCiudad Autónoma de Buenos AiresArgentinaSociedad Argentina de Investigaciones en Bioquímica y Biología MolecularUniversidad Nacional de Cuyo2013info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/244644Cloning of two new CDPK isoforms from Solanum tuberosum; Molecular mechanisms in cell signaling and gene expression; Ciudad Autónoma de Buenos Aires; Argentina; 2013; 33-330327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saib.org.ar/sites/default/files/BIOCELL-SAIB-2013.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:00:39Zoai:ri.conicet.gov.ar:11336/244644instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:00:39.483CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cloning of two new CDPK isoforms from Solanum tuberosum
title Cloning of two new CDPK isoforms from Solanum tuberosum
spellingShingle Cloning of two new CDPK isoforms from Solanum tuberosum
Fantino, Elisa Inés
CDPKs
CALCIO
SOLANUM TUBEROSUM
CHLOROPLASTS
title_short Cloning of two new CDPK isoforms from Solanum tuberosum
title_full Cloning of two new CDPK isoforms from Solanum tuberosum
title_fullStr Cloning of two new CDPK isoforms from Solanum tuberosum
title_full_unstemmed Cloning of two new CDPK isoforms from Solanum tuberosum
title_sort Cloning of two new CDPK isoforms from Solanum tuberosum
dc.creator.none.fl_str_mv Fantino, Elisa Inés
Santin, Franco
Ulloa, Rita Maria
author Fantino, Elisa Inés
author_facet Fantino, Elisa Inés
Santin, Franco
Ulloa, Rita Maria
author_role author
author2 Santin, Franco
Ulloa, Rita Maria
author2_role author
author
dc.subject.none.fl_str_mv CDPKs
CALCIO
SOLANUM TUBEROSUM
CHLOROPLASTS
topic CDPKs
CALCIO
SOLANUM TUBEROSUM
CHLOROPLASTS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Calcium-dependent protein kinases (CDPKs) comprise a multigene family of calcium sensors that play important roles regulating plant growth and development and plant responses to environmental stresses. To the moment six isoforms have been characterized in potato plants. In silico analysis of Solanum phureja genome allowed us to identify other 20 candidate genes encoding potential CDPK isoforms that cluster into four subgroups. Expression of 13 of these isoforms was detected in leaves and stolons from Solanum tuberosum cv Desirée plants. Using primers directed against their 5´and 3´utr sequences we amplified isoforms StCDPK25 and StCDPK17 belonging to subgroup 1 and 3 respectively. These fragments were cloned and sequenced. Isoform StCDPK25 shares 99% identity with the S. phureja coding sequence and presented all the characteristics of a CDPK. Cloning of StCDPK25 coding sequence in pDEST expression vector in order to obtain the recombinant enzyme tagged to 6xHis is currently in progress. Analysis of its sequence indicated that it could be targeted to chloroplasts; the N-terminal sequence will be fused to GFP and transient expression assays will be performed in Nicotiana benthamiana. On the contrary, only a 1 kb fragment containing exons 1, 2 and 7 was amplified for StCDPK17 suggesting that alternative splicing has occurred.
Fil: Fantino, Elisa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Santin, Franco. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ulloa, Rita Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Molecular mechanisms in cell signaling and gene expression
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
description Calcium-dependent protein kinases (CDPKs) comprise a multigene family of calcium sensors that play important roles regulating plant growth and development and plant responses to environmental stresses. To the moment six isoforms have been characterized in potato plants. In silico analysis of Solanum phureja genome allowed us to identify other 20 candidate genes encoding potential CDPK isoforms that cluster into four subgroups. Expression of 13 of these isoforms was detected in leaves and stolons from Solanum tuberosum cv Desirée plants. Using primers directed against their 5´and 3´utr sequences we amplified isoforms StCDPK25 and StCDPK17 belonging to subgroup 1 and 3 respectively. These fragments were cloned and sequenced. Isoform StCDPK25 shares 99% identity with the S. phureja coding sequence and presented all the characteristics of a CDPK. Cloning of StCDPK25 coding sequence in pDEST expression vector in order to obtain the recombinant enzyme tagged to 6xHis is currently in progress. Analysis of its sequence indicated that it could be targeted to chloroplasts; the N-terminal sequence will be fused to GFP and transient expression assays will be performed in Nicotiana benthamiana. On the contrary, only a 1 kb fragment containing exons 1, 2 and 7 was amplified for StCDPK17 suggesting that alternative splicing has occurred.
publishDate 2013
dc.date.none.fl_str_mv 2013
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/244644
Cloning of two new CDPK isoforms from Solanum tuberosum; Molecular mechanisms in cell signaling and gene expression; Ciudad Autónoma de Buenos Aires; Argentina; 2013; 33-33
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244644
identifier_str_mv Cloning of two new CDPK isoforms from Solanum tuberosum; Molecular mechanisms in cell signaling and gene expression; Ciudad Autónoma de Buenos Aires; Argentina; 2013; 33-33
0327-9545
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.saib.org.ar/sites/default/files/BIOCELL-SAIB-2013.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Universidad Nacional de Cuyo
publisher.none.fl_str_mv Universidad Nacional de Cuyo
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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