Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System

Autores
Metz, German Ernesto; Abeyá, María Mercedes; Serena, Maria Soledad; Panei, Carlos Javier; Diaz, Silvina; Echeverria, Maria Gabriela
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Our objective was to obtain Equine Arteritis Virus M protein in prokaryotic system to test it as immunogen. LP02/C Equine Arteritis Virus cDNA was used astemplate to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M wastransformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth comparedwith the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and differentinductors were analyzed but any of them showed an improvement in protein expression. Instead of E. coli M15strain, a new strain (E. coli BL21) was used andtransformed with the pQE30/M. This resolved in part the growth inhibition observed in E. coli M15 cells, but no the recovery yield of the protein. So, as all geneproducts that affect cells kinetics growth are considered to be toxic, we argue that the lower yields in M protein recovery could be attributed to an associated toxicityof EAV-M protein from LP02/C strain in this expression system.
Fil: Metz, German Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Abeyá, María Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Serena, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina
Fil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Materia
equine arteritis virus
LP02/C strain
M protein
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:11336/66956
Acceder
id CONICETDig_cab97ad78fe8e908bb59f415ec6e52c0
oai_identifier_str oai:ri.conicet.gov.ar:11336/66956
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 SystemMetz, German ErnestoAbeyá, María MercedesSerena, Maria SoledadPanei, Carlos JavierDiaz, SilvinaEcheverria, Maria Gabrielaequine arteritis virusLP02/C strainM proteinhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Our objective was to obtain Equine Arteritis Virus M protein in prokaryotic system to test it as immunogen. LP02/C Equine Arteritis Virus cDNA was used astemplate to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M wastransformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth comparedwith the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and differentinductors were analyzed but any of them showed an improvement in protein expression. Instead of E. coli M15strain, a new strain (E. coli BL21) was used andtransformed with the pQE30/M. This resolved in part the growth inhibition observed in E. coli M15 cells, but no the recovery yield of the protein. So, as all geneproducts that affect cells kinetics growth are considered to be toxic, we argue that the lower yields in M protein recovery could be attributed to an associated toxicityof EAV-M protein from LP02/C strain in this expression system.Fil: Metz, German Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Abeyá, María Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Serena, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; ArgentinaFil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaOpen Acces text2017-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/66956Metz, German Ernesto; Abeyá, María Mercedes; Serena, Maria Soledad; Panei, Carlos Javier; Diaz, Silvina; et al.; Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System; Open Acces text; Virology: Research and Reviews; 1; 2; 5-2017; 1-52514-4138CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.15761/VRR.1000111info:eu-repo/semantics/altIdentifier/url/https://www.oatext.com/Expression-of-M-Protein-from-LP02C-Equine-Arteritis-Virus-Inhibits-Growth-of-Escherichia-Coli-M15-Pqe30-System.phpinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-04-15T10:19:40Zoai:ri.conicet.gov.ar:11336/66956instacron: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:34982026-04-15 10:19:40.614CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
title Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
spellingShingle Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
Metz, German Ernesto
equine arteritis virus
LP02/C strain
M protein
title_short Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
title_full Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
title_fullStr Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
title_full_unstemmed Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
title_sort Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System
dc.creator.none.fl_str_mv Metz, German Ernesto
Abeyá, María Mercedes
Serena, Maria Soledad
Panei, Carlos Javier
Diaz, Silvina
Echeverria, Maria Gabriela
author Metz, German Ernesto
author_facet Metz, German Ernesto
Abeyá, María Mercedes
Serena, Maria Soledad
Panei, Carlos Javier
Diaz, Silvina
Echeverria, Maria Gabriela
author_role author
author2 Abeyá, María Mercedes
Serena, Maria Soledad
Panei, Carlos Javier
Diaz, Silvina
Echeverria, Maria Gabriela
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv equine arteritis virus
LP02/C strain
M protein
topic equine arteritis virus
LP02/C strain
M protein
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Our objective was to obtain Equine Arteritis Virus M protein in prokaryotic system to test it as immunogen. LP02/C Equine Arteritis Virus cDNA was used astemplate to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M wastransformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth comparedwith the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and differentinductors were analyzed but any of them showed an improvement in protein expression. Instead of E. coli M15strain, a new strain (E. coli BL21) was used andtransformed with the pQE30/M. This resolved in part the growth inhibition observed in E. coli M15 cells, but no the recovery yield of the protein. So, as all geneproducts that affect cells kinetics growth are considered to be toxic, we argue that the lower yields in M protein recovery could be attributed to an associated toxicityof EAV-M protein from LP02/C strain in this expression system.
Fil: Metz, German Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Abeyá, María Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Serena, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina
Fil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
description Our objective was to obtain Equine Arteritis Virus M protein in prokaryotic system to test it as immunogen. LP02/C Equine Arteritis Virus cDNA was used astemplate to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M wastransformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth comparedwith the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and differentinductors were analyzed but any of them showed an improvement in protein expression. Instead of E. coli M15strain, a new strain (E. coli BL21) was used andtransformed with the pQE30/M. This resolved in part the growth inhibition observed in E. coli M15 cells, but no the recovery yield of the protein. So, as all geneproducts that affect cells kinetics growth are considered to be toxic, we argue that the lower yields in M protein recovery could be attributed to an associated toxicityof EAV-M protein from LP02/C strain in this expression system.
publishDate 2017
dc.date.none.fl_str_mv 2017-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/66956
Metz, German Ernesto; Abeyá, María Mercedes; Serena, Maria Soledad; Panei, Carlos Javier; Diaz, Silvina; et al.; Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System; Open Acces text; Virology: Research and Reviews; 1; 2; 5-2017; 1-5
2514-4138
CONICET Digital
CONICET
url http://hdl.handle.net/11336/66956
identifier_str_mv Metz, German Ernesto; Abeyá, María Mercedes; Serena, Maria Soledad; Panei, Carlos Javier; Diaz, Silvina; et al.; Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System; Open Acces text; Virology: Research and Reviews; 1; 2; 5-2017; 1-5
2514-4138
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.15761/VRR.1000111
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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
application/pdf
application/pdf
application/pdf
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dc.publisher.none.fl_str_mv Open Acces text
publisher.none.fl_str_mv Open Acces text
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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