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

Autores
Metz, Germán Ernesto; Abeyá, María Mercedes; Serena, María Soledad; Panei, Carlos Javier; Díaz, Silvina; Echeverría, María 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 as template to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M was transformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth compared with the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and different inductors 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 and transformed 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 gene products 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 toxicity of EAV-M protein from LP02/C strain in this expression system.
Facultad de Ciencias Veterinarias (FCV)
Materia
Ciencias Veterinarias
equine arteritis virus
M protein
equinos
virus
Escherichia coli M15
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-nd/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/68168
Acceder
id SEDICI_3eb89a386e5fac37c6be13d6dd040724
oai_identifier_str oai:sedici.unlp.edu.ar:10915/68168
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 SystemMetz, Germán ErnestoAbeyá, María MercedesSerena, María SoledadPanei, Carlos JavierDíaz, SilvinaEcheverría, María GabrielaCiencias Veterinariasequine arteritis virusM proteinequinosvirusEscherichia coli M15Our 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 as template to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M was transformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth compared with the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and different inductors 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 and transformed 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 gene products 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 toxicity of EAV-M protein from LP02/C strain in this expression system.Facultad de Ciencias Veterinarias (FCV)2017-04-27info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://sedici.unlp.edu.ar/handle/10915/68168enginfo:eu-repo/semantics/altIdentifier/url/http://www.oatext.com/Expression-of-M-Protein-from-LP02C-Equine-Arteritis-Virus-Inhibits-Growth-of-Escherichia-Coli-M15-Pqe30-System.phpinfo:eu-repo/semantics/altIdentifier/issn/2514-4138info:eu-repo/semantics/altIdentifier/doi/10.15761/VRR.1000111info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/4.0/Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2026-04-15T11:18:07Zoai:sedici.unlp.edu.ar:10915/68168Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292026-04-15 11:18:07.884SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
title Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
spellingShingle Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
Metz, Germán Ernesto
Ciencias Veterinarias
equine arteritis virus
M protein
equinos
virus
Escherichia coli M15
title_short Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
title_full Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
title_fullStr Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
title_full_unstemmed Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
title_sort Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of <i>Escherichia Coli</i> M15-pQE30 System
dc.creator.none.fl_str_mv Metz, Germán Ernesto
Abeyá, María Mercedes
Serena, María Soledad
Panei, Carlos Javier
Díaz, Silvina
Echeverría, María Gabriela
author Metz, Germán Ernesto
author_facet Metz, Germán Ernesto
Abeyá, María Mercedes
Serena, María Soledad
Panei, Carlos Javier
Díaz, Silvina
Echeverría, María Gabriela
author_role author
author2 Abeyá, María Mercedes
Serena, María Soledad
Panei, Carlos Javier
Díaz, Silvina
Echeverría, María Gabriela
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Ciencias Veterinarias
equine arteritis virus
M protein
equinos
virus
Escherichia coli M15
topic Ciencias Veterinarias
equine arteritis virus
M protein
equinos
virus
Escherichia coli M15
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 as template to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M was transformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth compared with the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and different inductors 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 and transformed 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 gene products 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 toxicity of EAV-M protein from LP02/C strain in this expression system.
Facultad de Ciencias Veterinarias (FCV)
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 as template to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M was transformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth compared with the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and different inductors 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 and transformed 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 gene products 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 toxicity of EAV-M protein from LP02/C strain in this expression system.
publishDate 2017
dc.date.none.fl_str_mv 2017-04-27
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/68168
url http://sedici.unlp.edu.ar/handle/10915/68168
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.oatext.com/Expression-of-M-Protein-from-LP02C-Equine-Arteritis-Virus-Inhibits-Growth-of-Escherichia-Coli-M15-Pqe30-System.php
info:eu-repo/semantics/altIdentifier/issn/2514-4138
info:eu-repo/semantics/altIdentifier/doi/10.15761/VRR.1000111
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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