Taller De Micologia Avanzada
Páginas: 10 (2290 palabras)
Publicado: 17 de febrero de 2013
TALLER DE MICOLOGIA AVANZADA
PRESENTADO POR:
CARLOS DAVID ARANGO
PRESENTADO A:
M.S.C. CLAUDIA TAPIA LARIOS
UNIVERSIDAD COOPERATIVA DE COLOMBIA
FACULTAD CIENCIAS DE LA SALUD
PROGRAMA MEDICINA
GRUPO A
2012
Taller de Micología Avanzada
Medicina
Primer semestre
3.Lea el siguiente párrafo y responda las siguientes preguntas:
a.Cual es el hongo utilizado en este sistema biotecnológico. Defina sus características taxonómicas.
RESPUESTA:
LACCARIA BICOLOR
* Subdivisión: Basidiomycotina
* Clase: Homobasidiomycetes
* Subclase: Agaricomycetidae
* Orden: Tricholomatales
* Familia: Tricholomataceae
b. Porque este hongo resulto ser el mejor modelo genómico a nivel para integración de plásmidos.RESPUESTA:
hecho que este hongo fue transformado vía AMT lo hizo un excelente modelo para estudios funcionales genómicos
c. Que es una trasformación AMP y cual su uso primario en Lacaria bicolor.
RESPUESTA:
AMP: debido a que no existen datos sobre el patrón de integración en el genoma de laccaria disponibles hemos optimizado un método de rescate plasmido para este hongo. Para estefin, el vector de transformación (PHG/ p BSK) se construyo permitiendo el rescate de los t DNA adecuadas uniones de ADN fronteragenomicas en E. coli.
El método de rescate de plasmido se puede utilizar para la resolución de T- DNA en los sitios de integración laccaria.
POST-GENOMIC STUDIES IN MYCORRHIZA
Kemppainen M1, Duplessis S, Martin F & Pardo AG
Departamento de Ciencia yTecnología, Universidad
Nacional de Quilmes. B1876BXD Bernal, Provincia de
Buenos Aires, Argentina. INRA-Nancy 54280 Champenoux,
France. E-mail: apardo@unq.edu.ar .
Agrobacterium-mediated gene transfer (AMT) in fungi is currently opening a new era for fungal genetics. As whole genome sequences of several fungi are being released studies about T-DNA integration patterns and the development offunctional genomic tools are urgently needed. The first genome sequence of a symbiotic fungus, the basidiomycete Laccaria bicolor, became public in July 2006 (http://genome.jgi-psf.org/Lacbi1/Lacbi1.home.html). Now when Laccaria genome is available and the fact that this fungus was transformed via AMT (Kemppainen et al., 2005) makes it an excellent model for functional genomic studies
in mycorrhizalresearch. Because no data on the integration pattern in Laccaria genome were available we optimized a plasmid rescue approach for this fungus. To this end the transformation vector (pHg/pBSk) was constructed allowing the rescue of the TDNA right border-genomic DNA junctions in E.coli.
A total of 51 Agrobacterium - transformed fungal lines were analyzed. Sixty nine percent were successfullyrescued for the right border of which 87% were resolved for genomic integration sequences. Our results demonstrate that the plasmid rescue approach can be used for resolving T-DNA integration sites in Laccaria. The RB was well conserved during transformation of this fungus and the integration analysis showed no clear sequence homology between different genomic sites. Neither obvious sequencesimilarities were found between these sites and the T-strand borders indicating nonhomologous and random integration of the 203 transgenes. Majority (74%) of the integrations were located in predicted genes. Three fungal lines with integrations in putative ORFs were subjected to expression analysis by RT-PCR. Our results demonstrate that AMT can be used for gene
validation in Laccaria. On the otherhand, we have developed the RNAi technology in Laccaria in order to achieve the knock-down of selected fungal genes and targeted GFP expression. The availability of these functional genomic tools will certainly help in the development of the post-genomic era in mycorrhizal research.
4. Investigue cuales son las enfermedades nosocomiales mas frecuentes de origen fúngico, cuales son sus...
PRESENTADO POR:
CARLOS DAVID ARANGO
PRESENTADO A:
M.S.C. CLAUDIA TAPIA LARIOS
UNIVERSIDAD COOPERATIVA DE COLOMBIA
FACULTAD CIENCIAS DE LA SALUD
PROGRAMA MEDICINA
GRUPO A
2012
Taller de Micología Avanzada
Medicina
Primer semestre
3.Lea el siguiente párrafo y responda las siguientes preguntas:
a.Cual es el hongo utilizado en este sistema biotecnológico. Defina sus características taxonómicas.
RESPUESTA:
LACCARIA BICOLOR
* Subdivisión: Basidiomycotina
* Clase: Homobasidiomycetes
* Subclase: Agaricomycetidae
* Orden: Tricholomatales
* Familia: Tricholomataceae
b. Porque este hongo resulto ser el mejor modelo genómico a nivel para integración de plásmidos.RESPUESTA:
hecho que este hongo fue transformado vía AMT lo hizo un excelente modelo para estudios funcionales genómicos
c. Que es una trasformación AMP y cual su uso primario en Lacaria bicolor.
RESPUESTA:
AMP: debido a que no existen datos sobre el patrón de integración en el genoma de laccaria disponibles hemos optimizado un método de rescate plasmido para este hongo. Para estefin, el vector de transformación (PHG/ p BSK) se construyo permitiendo el rescate de los t DNA adecuadas uniones de ADN fronteragenomicas en E. coli.
El método de rescate de plasmido se puede utilizar para la resolución de T- DNA en los sitios de integración laccaria.
POST-GENOMIC STUDIES IN MYCORRHIZA
Kemppainen M1, Duplessis S, Martin F & Pardo AG
Departamento de Ciencia yTecnología, Universidad
Nacional de Quilmes. B1876BXD Bernal, Provincia de
Buenos Aires, Argentina. INRA-Nancy 54280 Champenoux,
France. E-mail: apardo@unq.edu.ar .
Agrobacterium-mediated gene transfer (AMT) in fungi is currently opening a new era for fungal genetics. As whole genome sequences of several fungi are being released studies about T-DNA integration patterns and the development offunctional genomic tools are urgently needed. The first genome sequence of a symbiotic fungus, the basidiomycete Laccaria bicolor, became public in July 2006 (http://genome.jgi-psf.org/Lacbi1/Lacbi1.home.html). Now when Laccaria genome is available and the fact that this fungus was transformed via AMT (Kemppainen et al., 2005) makes it an excellent model for functional genomic studies
in mycorrhizalresearch. Because no data on the integration pattern in Laccaria genome were available we optimized a plasmid rescue approach for this fungus. To this end the transformation vector (pHg/pBSk) was constructed allowing the rescue of the TDNA right border-genomic DNA junctions in E.coli.
A total of 51 Agrobacterium - transformed fungal lines were analyzed. Sixty nine percent were successfullyrescued for the right border of which 87% were resolved for genomic integration sequences. Our results demonstrate that the plasmid rescue approach can be used for resolving T-DNA integration sites in Laccaria. The RB was well conserved during transformation of this fungus and the integration analysis showed no clear sequence homology between different genomic sites. Neither obvious sequencesimilarities were found between these sites and the T-strand borders indicating nonhomologous and random integration of the 203 transgenes. Majority (74%) of the integrations were located in predicted genes. Three fungal lines with integrations in putative ORFs were subjected to expression analysis by RT-PCR. Our results demonstrate that AMT can be used for gene
validation in Laccaria. On the otherhand, we have developed the RNAi technology in Laccaria in order to achieve the knock-down of selected fungal genes and targeted GFP expression. The availability of these functional genomic tools will certainly help in the development of the post-genomic era in mycorrhizal research.
4. Investigue cuales son las enfermedades nosocomiales mas frecuentes de origen fúngico, cuales son sus...
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