Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies
Background: Snakin-1 (StSN1) is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanum tuberosum. Its biotechnological potential has been already recognized since it exhibits in vivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologous peptides, in...
| Autores principales: | , , , , , , |
|---|---|
| Formato: | info:eu-repo/semantics/article |
| Lenguaje: | Inglés |
| Publicado: |
2018
|
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/1964 https://bmcbiotechnol.biomedcentral.com/track/pdf/10.1186/s12896-017-0401-2?site=bmcbiotechnol.biomedcentral.com https://doi.org/10.1186/s12896-017-0401-2 |
| _version_ | 1855034881923350528 |
|---|---|
| author | Almasia, Natalia Ines Molinari, Maria Paula Maroniche, Guillermo Andrés Nahirñak, Vanesa Barrios Baron, Maria Pilar Taboga, Oscar Alberto Vazquez Rovere, Cecilia |
| author_browse | Almasia, Natalia Ines Barrios Baron, Maria Pilar Maroniche, Guillermo Andrés Molinari, Maria Paula Nahirñak, Vanesa Taboga, Oscar Alberto Vazquez Rovere, Cecilia |
| author_facet | Almasia, Natalia Ines Molinari, Maria Paula Maroniche, Guillermo Andrés Nahirñak, Vanesa Barrios Baron, Maria Pilar Taboga, Oscar Alberto Vazquez Rovere, Cecilia |
| author_sort | Almasia, Natalia Ines |
| collection | INTA Digital |
| description | Background: Snakin-1 (StSN1) is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanum tuberosum. Its biotechnological potential has been already recognized since it exhibits in vivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologous peptides, in a soluble native state using bacterial, yeast or synthetic expression systems have presented production bottlenecks such as insolubility, misfolding or low yields.
Results: In this work, we successfully expressed a recombinant StSN1 (rSN1) in Spodoptera frugiperda (Sf9) insect cells by optimizing several of the parameters for its expression in the baculovirus expression system. The recombinant
peptide lacking its putative signal peptide was soluble and was present in the nuclear fraction of infected Sf9 cells. An optimized purification procedure allowed the production of rSN1 that was used for immunization of mice, which gave rise to polyclonal antibodies that detect the native protein in tissue extracts of both agroinfiltrated plants and stable transgenic lines. Our results demonstrated that this system circumvents all the difficulties associated with recombinant antimicrobial peptides expression in other heterologous systems. |
| format | info:eu-repo/semantics/article |
| id | INTA1964 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| record_format | dspace |
| spelling | INTA19642019-10-23T18:09:19Z Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies Almasia, Natalia Ines Molinari, Maria Paula Maroniche, Guillermo Andrés Nahirñak, Vanesa Barrios Baron, Maria Pilar Taboga, Oscar Alberto Vazquez Rovere, Cecilia Papa Péptidos Antimicrobianos Baculovirus Anticuerpos Potatoes Peptides Antimicrobials Antibodies Background: Snakin-1 (StSN1) is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanum tuberosum. Its biotechnological potential has been already recognized since it exhibits in vivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologous peptides, in a soluble native state using bacterial, yeast or synthetic expression systems have presented production bottlenecks such as insolubility, misfolding or low yields. Results: In this work, we successfully expressed a recombinant StSN1 (rSN1) in Spodoptera frugiperda (Sf9) insect cells by optimizing several of the parameters for its expression in the baculovirus expression system. The recombinant peptide lacking its putative signal peptide was soluble and was present in the nuclear fraction of infected Sf9 cells. An optimized purification procedure allowed the production of rSN1 that was used for immunization of mice, which gave rise to polyclonal antibodies that detect the native protein in tissue extracts of both agroinfiltrated plants and stable transgenic lines. Our results demonstrated that this system circumvents all the difficulties associated with recombinant antimicrobial peptides expression in other heterologous systems. Inst. de Biotecnología Fil: Almasia, Natalia Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Molinari, Maria Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Maroniche, Guillermo Andrés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina Fil: Nahirñak, Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Barrios Baron, Maria Pilar. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vazquez Rovere, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Labintex-Instituto Nacional de Tecnología Agropecuaria (INTA). Francia 2018-03-05T17:08:25Z 2018-03-05T17:08:25Z 2017 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/1964 https://bmcbiotechnol.biomedcentral.com/track/pdf/10.1186/s12896-017-0401-2?site=bmcbiotechnol.biomedcentral.com 1472-6750 https://doi.org/10.1186/s12896-017-0401-2 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf BMC Biotechnology 17 : 75 (2017) |
| spellingShingle | Papa Péptidos Antimicrobianos Baculovirus Anticuerpos Potatoes Peptides Antimicrobials Antibodies Almasia, Natalia Ines Molinari, Maria Paula Maroniche, Guillermo Andrés Nahirñak, Vanesa Barrios Baron, Maria Pilar Taboga, Oscar Alberto Vazquez Rovere, Cecilia Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title | Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title_full | Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title_fullStr | Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title_full_unstemmed | Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title_short | Successful production of the potato antimicrobial peptide Snakin-1 in baculovirus-infected insect cells and development of specific antibodies |
| title_sort | successful production of the potato antimicrobial peptide snakin 1 in baculovirus infected insect cells and development of specific antibodies |
| topic | Papa Péptidos Antimicrobianos Baculovirus Anticuerpos Potatoes Peptides Antimicrobials Antibodies |
| url | http://hdl.handle.net/20.500.12123/1964 https://bmcbiotechnol.biomedcentral.com/track/pdf/10.1186/s12896-017-0401-2?site=bmcbiotechnol.biomedcentral.com https://doi.org/10.1186/s12896-017-0401-2 |
| work_keys_str_mv | AT almasianataliaines successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT molinarimariapaula successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT maronicheguillermoandres successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT nahirnakvanesa successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT barriosbaronmariapilar successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT tabogaoscaralberto successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies AT vazquezroverececilia successfulproductionofthepotatoantimicrobialpeptidesnakin1inbaculovirusinfectedinsectcellsanddevelopmentofspecificantibodies |