Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
- Autores
- Rodríguez Durán, Jessica Jenireth; Gomez, Karina Andrea; Potenza, Mariana
- Año de publicación
- 2019
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi.
Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimenta
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio - Materia
-
Trypanosoma cruzi
Calcium binding
Parasitology - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/201594
Ver los metadatos del registro completo
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Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruziRodríguez Durán, Jessica JenirethGomez, Karina AndreaPotenza, MarianaTrypanosoma cruziCalcium bindingParasitologyhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi.Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaLXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de Investigación ClínicaAsociación Argentina de Farmacología ExperimentaSociedad Argentina de BiologíaSociedad Argentina de ProtozoologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioFundación Revista MedicinaCostas, Monica AlejandraMarino, Gabriela InésAzurmendi, Pablo Javier2019info: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/11336/201594Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-1210025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol79-19/s4/vol79_s4.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 Técnicas2025-10-15T15:03:46Zoai:ri.conicet.gov.ar:11336/201594instacron: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:34982025-10-15 15:03:46.562CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
title |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
spellingShingle |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi Rodríguez Durán, Jessica Jenireth Trypanosoma cruzi Calcium binding Parasitology |
title_short |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
title_full |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
title_fullStr |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
title_full_unstemmed |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
title_sort |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi |
dc.creator.none.fl_str_mv |
Rodríguez Durán, Jessica Jenireth Gomez, Karina Andrea Potenza, Mariana |
author |
Rodríguez Durán, Jessica Jenireth |
author_facet |
Rodríguez Durán, Jessica Jenireth Gomez, Karina Andrea Potenza, Mariana |
author_role |
author |
author2 |
Gomez, Karina Andrea Potenza, Mariana |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Costas, Monica Alejandra Marino, Gabriela Inés Azurmendi, Pablo Javier |
dc.subject.none.fl_str_mv |
Trypanosoma cruzi Calcium binding Parasitology |
topic |
Trypanosoma cruzi Calcium binding Parasitology |
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 homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi. Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio Mar del Plata Argentina Sociedad Argentina de Investigación Clínica Asociación Argentina de Farmacología Experimenta Sociedad Argentina de Biología Sociedad Argentina de Protozoología Asociación Argentina de Nanomedicinas Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio |
description |
Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019 |
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/11336/201594 Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-121 0025-7680 1669-9106 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/201594 |
identifier_str_mv |
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-121 0025-7680 1669-9106 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://medicinabuenosaires.com/revistas/vol79-19/s4/vol79_s4.pdf |
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info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
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application/pdf application/pdf application/pdf |
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Internacional |
dc.publisher.none.fl_str_mv |
Fundación Revista Medicina |
publisher.none.fl_str_mv |
Fundación Revista Medicina |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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