Projecto: “Vacuna derivada de plantas para la prevención de peste bubónica y neumónica”
Lugar de trabajo: Centro de Enfermedades Infecciosas y Vacunas (CIDV)- Biodesign A - Arizona State University, Tempe, Arizona, U.S.A. (2003 – presente).
Cargo: Investigadora Post-doctoral
Lugar de trabajo: Centro de Enfermedades Infecciosas y Vacunas (CIDV)- Biodesign A - Arizona State University, Tempe, Arizona, U.S.A. (2003 – presente).
Cargo: Investigadora Post-doctoral
El objetivo de este proyecto fue expresar antígenos de la bacteria Yersinia pestis en tomate para desarrollar una vacuna oral derivada de plantas para prevenir peste bubónica y neumónica. Obtuvimos tomate transgénico expresando altos niveles de proteina de fusion F1-V (F1 y V son los dos antígenos más inmunogénicos de Y. pestis. El tomate conteniendo la vacuna fue disecado y convertido en polvo con el objeto de concentrar el antigeno F1-V y obtener una vacuna estable a temperatura ambiente. La vacuna derivada del tomate fue capaz de inducir una respuesta inmune satisfactoria en ratones. Los resultados obtenidos fueron publicados (Alvarez et al., 2004; Alvarez et al., 2006) y presentados en conferencias nacionales e internacionales.
La Dra. Alvarez diseñó y llevó a cabo todos los experimentos de este proyecto incluyendo la construcción de los plásmidos usados en la transformación, la selección y caracterización de las plantas transgénicas y los ensayos en animales para testear la vacuna derivada del tomate.
Projecto: “Reversión del silenciamiento génico de f1-v inducida por la expresión transiente y estable de TBSV-P19 en tomato”
Lugar de trabajo: Lugar de trabajo: Centro de Enfermedades Infecciosas y Vacunas (CIDV)- Biodesign A - Arizona State University, Tempe, Arizona, U.S.A. (2003- present).
La Dra. Alvarez diseñó y llevó a cabo todos los experimentos de este proyecto incluyendo la construcción de los plásmidos usados en la transformación, la selección y caracterización de las plantas transgénicas y los ensayos en animales para testear la vacuna derivada del tomate.
Projecto: “Reversión del silenciamiento génico de f1-v inducida por la expresión transiente y estable de TBSV-P19 en tomato”
Lugar de trabajo: Lugar de trabajo: Centro de Enfermedades Infecciosas y Vacunas (CIDV)- Biodesign A - Arizona State University, Tempe, Arizona, U.S.A. (2003- present).
Cargo: Investigadora Post-doctoral
El objetivo de este proyecto fue restaurar la expresión del gen f1-v en plantas de tomate silenciadas. A pesar de que estas plantas contenian un alto numero de copias del gen f1-v, los niveles de proteina F1-V fueron muy bajos o indetectables en hojas y frutos. Para recuperar la expresión del gen f1-v, super-transformamos las plantas silenciadas con el gen p19, un supresor de silenciamiento génico proveniente del virus del tomate TBSV. Las plantas que acumularon los mayores niveles de proteína P19 en transiente o en estable fueron también las que tuvieron el mayor contenido de proteina F1-V en hojas y frutos. Los niveles de proteinas F1-V en fruto de plantas con reversión de silenciamiento fueron significativamente mayores que en las plantas consideradas "F1-V elite" no silenciadas. Estos resultados confirman el potencial del uso de P19 para aumentar la acumulación de proteinas de interes en plantas transgenicas (Alvarez et al., manuscrito en revision enviado recientemente a la revista Plant Molecular Biology).
Dr. Alvarez diseñó y llevó a cabo todos los experimentos de este proyecto incluyendo la transformacion de tomate y la caracterizacion molecular de las plantas transgenicas obtenidas.
El objetivo de este proyecto fue restaurar la expresión del gen f1-v en plantas de tomate silenciadas. A pesar de que estas plantas contenian un alto numero de copias del gen f1-v, los niveles de proteina F1-V fueron muy bajos o indetectables en hojas y frutos. Para recuperar la expresión del gen f1-v, super-transformamos las plantas silenciadas con el gen p19, un supresor de silenciamiento génico proveniente del virus del tomate TBSV. Las plantas que acumularon los mayores niveles de proteína P19 en transiente o en estable fueron también las que tuvieron el mayor contenido de proteina F1-V en hojas y frutos. Los niveles de proteinas F1-V en fruto de plantas con reversión de silenciamiento fueron significativamente mayores que en las plantas consideradas "F1-V elite" no silenciadas. Estos resultados confirman el potencial del uso de P19 para aumentar la acumulación de proteinas de interes en plantas transgenicas (Alvarez et al., manuscrito en revision enviado recientemente a la revista Plant Molecular Biology).
Dr. Alvarez diseñó y llevó a cabo todos los experimentos de este proyecto incluyendo la transformacion de tomate y la caracterizacion molecular de las plantas transgenicas obtenidas.
Projecto: “Vacuna anticonceptiva derivada de plantas para uso en control de poblaciones de animales”
Lugar de trabajo: School of Life Sciences, Arizona State University, Tempe, Arizona, U.S.A. (2001 – 2002).
Cargos: Investigadora visitante y becaria de la Fundación Antorchas (Argentina).
The common brushtail possum is a marsupial native from Australia which was introduced to New Zealand by Europeans to establish a fur industry. They soon escaped into the wild where they have thrived as an invasive species with around 60 million individuals estimated. There have been numerous attempts to eradicate them because of the damage they do to native trees and wildlife, as well as acting as a carrier of bovine tuberculosis. The goal of this project was to develop an oral plant-derived vaccine to be used as immuno-contraceptive specie specific for possum population control. The seven amino acid epitope from ZP3 (zona pellucida glycoprotein 3) was fused to the heat-labile toxin (LT) of enterotoxigenic Escherichia coli and expressed in tomato. The fusion protein was found to assemble into pentamers and had an average expression level of 37.8 μg/g in freeze-dried transgenic tissues. The species-specific nature of this epitope was shown by the inability of antibodies raised against non-target species to detect the LTB fusion protein (Walmsley et al., 2003).
Dr Alvarez collaborated on this project doing the molecular characterization of the transgenic tomato plants expressing the fusion protein LTB-ZP3.
Project0: “Analisis de la funcionalidad de la masa derivada de harina de trigo transgénico”
Lugar de trabajo: Escuela Tecnica Superior de Ingenieros Agronomos (ETSIA), Departmento de Genetica y Mejoramiento de Plantas, Universidad Politecnica de Madrid (UPM), Madrid, ESPANA (2000)
Cargo: Research Scholar and Fellow of British Council and Fundacion Antorchas (Argentina).
The goal of this project was to determine the new physical properties of the dough made with flour from the different transgenic wheat lines expressing alleles of glutenins associated with better bread-making quality. The rheological properties of flours from five different lines of transgenic wheats that either express or over-express subunits 1Dx5 or 1Ax1 were analyzed by mixograph assays and SDS sedimentation tests. In one of the transgenic lines, the over-expression of subunit 1Dx5 resulted in a 2-fold increase in mixing time, associated with a significant improvement in dough strength, and a lower resistance breakdown, suggesting an important increase in dough quality (Alvarez et. al., 2001a; Alvarez et al., 2001 b).
Dr. Alvarez designed all the experiments and pursed all the dough analysis with the collaboration of colleagues at the Polytechnic University of Madrid, Spain.
Projecto (Tesis Doctoral): “Mejoramiento de la calidad nutricional y panadera del trigo por ingenieria genética".
Lugar de trabajo:
1) Centro de Estudios Fotosinteticos y Bioquimicos (CEFOBI), Universidad Nacional de Rosario, ARGENTINA (1995 – 2000)
2) Cell Biology Department, Institute of Arable Crops Research (IACR), Long Ashton Research Station, Bristol University, Bristol, INGLATERRA (2000)
Cargos:
1) Estudiante de doctorado y becaria del CONICET (Comision Nacional de Investigaciones Cientificas y Tecnicas) (1995-2000)
2) Cientifica visitante y becaria de British Council/ Fundación Antorchas (Argentina) (2000)
The main goal of this project was to improve the bread-making quality of wheat expressing high molecular weight (HMW) subunits of glutenins associated with better dough quality for bread.
Wheat HMW glutenin subunit genes 1Ax1 and 1Dx5 were introduced and either expressed or over-expressed into a commercial wheat cultivar that already expresses five subunits. Six independent transgenic wheat lines were obtained and characterized by SDS-PAGE and Southern-blot analyses. The 1Dx5 gene was over-expressed in two lines without changes in the other endosperm proteins. Two wheat lines expressed the 1Ax1 transgene with associated silencing of the 1Ax2* endogenous subunit. Southern analysis of the four events confirmed transformation and suggested that the transgenes were present in low gene copy number. Silencing of all the HMW glutenin subunits was observed in two different transgenic wheat lines expressing the 1Ax1 subunit transgene and over-expressing the 1Dx5 gene. Transgenes and expression patterns were stably transmitted to the progenies in all the events except one where in some of the segregating T2 seeds the silencing of all HMW glutenin subunits was reverted associated with a drastic loss of transgenes from a high to a low copy number.
This project was part of Dr. Alvarez's Ph.D Thesis and the results obtained were published in different journals (Alvarez et al., 2000; Alvarez et al., 2001).
Projecto: “Estudio de la presencia de fimbrias P en bacterias causantes de infección urinaria”.
Lugar de trabajo: Departmento de Microbiologia, Facultad de Bioquimica, Universidad Nacional de Rosario, ARGENTINA (1992-1993)
Cargo: Estudiante de Bioquimica y becaria de la fundacion de la Universidad Nacional de Rosario.
The long term goal of this project was to study the presence of glitter cells in the urinary sediment as a marker of high urinary infection by Enterobacterias mannose resistant that express fimbriae Pap. A total of 143 Enterobactereaceae strains isolated from patients with urinary infection were classified by biochemical and serological tests. The adhesive ability of the bacteria was determined using the human red cells group A agglutination test. The erythrocytes were suspended in phosphate buffer saline (PBS) either with or without mannose for testing mannose-resistant (MR-HA) or mannose- sensitive (MS-HA) hemagglutination. The number of Escherichia coli strains that expressed mannose-resistant fimbriae was 2-fold higher than the strains with mannose-sensitive fimbriae. There was a correlation between high urinary infections produced by E. coli strains with mannose-resistant fimbriae and presence of cells with special characteristics, glitter cells, in the urinary sediment.
Dr. Alvarez isolated and characterized the bacteria causing urinary infections using biochemical and serological tests. She determined the presence of mannose-resistant fimbriae in the bacteria and detected the glitter cells in the urinary sediment (Alvarez et al., 1994).
Lugar de trabajo: School of Life Sciences, Arizona State University, Tempe, Arizona, U.S.A. (2001 – 2002).
Cargos: Investigadora visitante y becaria de la Fundación Antorchas (Argentina).
The common brushtail possum is a marsupial native from Australia which was introduced to New Zealand by Europeans to establish a fur industry. They soon escaped into the wild where they have thrived as an invasive species with around 60 million individuals estimated. There have been numerous attempts to eradicate them because of the damage they do to native trees and wildlife, as well as acting as a carrier of bovine tuberculosis. The goal of this project was to develop an oral plant-derived vaccine to be used as immuno-contraceptive specie specific for possum population control. The seven amino acid epitope from ZP3 (zona pellucida glycoprotein 3) was fused to the heat-labile toxin (LT) of enterotoxigenic Escherichia coli and expressed in tomato. The fusion protein was found to assemble into pentamers and had an average expression level of 37.8 μg/g in freeze-dried transgenic tissues. The species-specific nature of this epitope was shown by the inability of antibodies raised against non-target species to detect the LTB fusion protein (Walmsley et al., 2003).
Dr Alvarez collaborated on this project doing the molecular characterization of the transgenic tomato plants expressing the fusion protein LTB-ZP3.
Project0: “Analisis de la funcionalidad de la masa derivada de harina de trigo transgénico”
Lugar de trabajo: Escuela Tecnica Superior de Ingenieros Agronomos (ETSIA), Departmento de Genetica y Mejoramiento de Plantas, Universidad Politecnica de Madrid (UPM), Madrid, ESPANA (2000)
Cargo: Research Scholar and Fellow of British Council and Fundacion Antorchas (Argentina).
The goal of this project was to determine the new physical properties of the dough made with flour from the different transgenic wheat lines expressing alleles of glutenins associated with better bread-making quality. The rheological properties of flours from five different lines of transgenic wheats that either express or over-express subunits 1Dx5 or 1Ax1 were analyzed by mixograph assays and SDS sedimentation tests. In one of the transgenic lines, the over-expression of subunit 1Dx5 resulted in a 2-fold increase in mixing time, associated with a significant improvement in dough strength, and a lower resistance breakdown, suggesting an important increase in dough quality (Alvarez et. al., 2001a; Alvarez et al., 2001 b).
Dr. Alvarez designed all the experiments and pursed all the dough analysis with the collaboration of colleagues at the Polytechnic University of Madrid, Spain.
Projecto (Tesis Doctoral): “Mejoramiento de la calidad nutricional y panadera del trigo por ingenieria genética".
Lugar de trabajo:
1) Centro de Estudios Fotosinteticos y Bioquimicos (CEFOBI), Universidad Nacional de Rosario, ARGENTINA (1995 – 2000)
2) Cell Biology Department, Institute of Arable Crops Research (IACR), Long Ashton Research Station, Bristol University, Bristol, INGLATERRA (2000)
Cargos:
1) Estudiante de doctorado y becaria del CONICET (Comision Nacional de Investigaciones Cientificas y Tecnicas) (1995-2000)
2) Cientifica visitante y becaria de British Council/ Fundación Antorchas (Argentina) (2000)
The main goal of this project was to improve the bread-making quality of wheat expressing high molecular weight (HMW) subunits of glutenins associated with better dough quality for bread.
Wheat HMW glutenin subunit genes 1Ax1 and 1Dx5 were introduced and either expressed or over-expressed into a commercial wheat cultivar that already expresses five subunits. Six independent transgenic wheat lines were obtained and characterized by SDS-PAGE and Southern-blot analyses. The 1Dx5 gene was over-expressed in two lines without changes in the other endosperm proteins. Two wheat lines expressed the 1Ax1 transgene with associated silencing of the 1Ax2* endogenous subunit. Southern analysis of the four events confirmed transformation and suggested that the transgenes were present in low gene copy number. Silencing of all the HMW glutenin subunits was observed in two different transgenic wheat lines expressing the 1Ax1 subunit transgene and over-expressing the 1Dx5 gene. Transgenes and expression patterns were stably transmitted to the progenies in all the events except one where in some of the segregating T2 seeds the silencing of all HMW glutenin subunits was reverted associated with a drastic loss of transgenes from a high to a low copy number.
This project was part of Dr. Alvarez's Ph.D Thesis and the results obtained were published in different journals (Alvarez et al., 2000; Alvarez et al., 2001).
Projecto: “Estudio de la presencia de fimbrias P en bacterias causantes de infección urinaria”.
Lugar de trabajo: Departmento de Microbiologia, Facultad de Bioquimica, Universidad Nacional de Rosario, ARGENTINA (1992-1993)
Cargo: Estudiante de Bioquimica y becaria de la fundacion de la Universidad Nacional de Rosario.
The long term goal of this project was to study the presence of glitter cells in the urinary sediment as a marker of high urinary infection by Enterobacterias mannose resistant that express fimbriae Pap. A total of 143 Enterobactereaceae strains isolated from patients with urinary infection were classified by biochemical and serological tests. The adhesive ability of the bacteria was determined using the human red cells group A agglutination test. The erythrocytes were suspended in phosphate buffer saline (PBS) either with or without mannose for testing mannose-resistant (MR-HA) or mannose- sensitive (MS-HA) hemagglutination. The number of Escherichia coli strains that expressed mannose-resistant fimbriae was 2-fold higher than the strains with mannose-sensitive fimbriae. There was a correlation between high urinary infections produced by E. coli strains with mannose-resistant fimbriae and presence of cells with special characteristics, glitter cells, in the urinary sediment.
Dr. Alvarez isolated and characterized the bacteria causing urinary infections using biochemical and serological tests. She determined the presence of mannose-resistant fimbriae in the bacteria and detected the glitter cells in the urinary sediment (Alvarez et al., 1994).
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