Caracterización bioquímica de la proteína AC29 de Alexandrium catenella
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Fecha
1999
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Profesor/a Guía
Facultad/escuela
Idioma
es
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Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
En Chile la mayoría de los estudios relacionados con el dinoflagelado tóxico
Alexandrium catenella, productor del Veneno Paralizante de los Mariscos (VPM),
han sido orientados a evaluar su presencia en el mar mediante los programas de
monitoreo fitoplanctónico, detección del VPM en los mariscos y sus perfiles
toxicológicos, especialmente de aquellos moluscos que son consumidos por el
hombre, sin embargo, existen pocos estudios sobre la biología del organismo.
El objetivo esta tesis fue desarrollar un modelo para estudiar la expresión
de proteínas en A. catenella. Se observó una banda proteica de una masa
molecular de 29 kDa mediante un análisis electroforético de proteínas totales de
cepas chilenas de A. catenella y diferentes especies del género Alexandrium. Se
determinó mediante análisis electroforético en dos dimensiones y Western blot
que esta banda está compuesta por al menos 6 proteínas de similar masa
molecular pero con diferente punto isoeléctrico, donde una de ellas es mas
abundante y fue denominada AC29.
La proteína AC29 fue inmunolocalizada mediante microscopía electrónica
en los cloroplastos y asociada a las membranas tilacoidales. Cuando las células
fueron cultivadas en condiciones limitantes de luz, la expresión de la proteína
disminuye, sugiriendo que la expresión de AC29 es regulada por la luz. La
expresión de AC29 está restringida a algunas especies fitoplanctónicas y no es
expresada en espinaca. Finalmente se obtuvo la secuencia del amino terminal de
AC29.
In Chile, most of the studies related to the toxic dinoflagellate, Alexandrium catenella, producer of Paralytic Shellfish Poisoning (PSP), have been oriented to demonstrate through phytoplanktonic monitoring programs its presence in the sea, or to PSP detection and its profile in shellfish commercialized for human consumption. However, there are few studies on the biology of this organism. The aim of this thesis was to develop a model to study protein expression in A. catenella. A major protein band was observed with a molecular mass of 29 kDa by electrophoretic analysis of extracts of chilean Alexandrium catenella clones and of different Alexandrium species. This 29 kDa band is composed by at least 6 different proteins of similar molecular mass, but with different isoelectric points as shown by two dimensional electrophoretic analysis followed by Western blot. This analysis also showed the presence of an abundant protein that was denominated AC29. The AC29 protein was located within the chloroplast by immunoelectronmicroscopy and associated to the lamella membrane. When cells in culture were light-deprived, protein expression decreased, suggesting that AC29 gene expression is regulated by light. The expression of AC29 is restricted to phytoplanktonic species and is not present in higher plants such as spinach. Finally, the amino terminal sequence of AC29 was obtained.
In Chile, most of the studies related to the toxic dinoflagellate, Alexandrium catenella, producer of Paralytic Shellfish Poisoning (PSP), have been oriented to demonstrate through phytoplanktonic monitoring programs its presence in the sea, or to PSP detection and its profile in shellfish commercialized for human consumption. However, there are few studies on the biology of this organism. The aim of this thesis was to develop a model to study protein expression in A. catenella. A major protein band was observed with a molecular mass of 29 kDa by electrophoretic analysis of extracts of chilean Alexandrium catenella clones and of different Alexandrium species. This 29 kDa band is composed by at least 6 different proteins of similar molecular mass, but with different isoelectric points as shown by two dimensional electrophoretic analysis followed by Western blot. This analysis also showed the presence of an abundant protein that was denominated AC29. The AC29 protein was located within the chloroplast by immunoelectronmicroscopy and associated to the lamella membrane. When cells in culture were light-deprived, protein expression decreased, suggesting that AC29 gene expression is regulated by light. The expression of AC29 is restricted to phytoplanktonic species and is not present in higher plants such as spinach. Finally, the amino terminal sequence of AC29 was obtained.
Notas
Tesis (Ingeniero en Acuicultura, Licenciado en Ciencias de la Aquicultura)
Palabras clave
Dinoflagelados, Toxinas Marinas