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2014, Número 1

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Biotecnol Apl 2014; 31 (1)


Aislamiento y clonaje del gen de la proteína antiviral PAP-I de Phytolacca americana

Mahfouze HA, El-Dougdoug KA, Othman BA, Gomaa MA

Idioma: Ingles.
Referencias bibliográficas: 24
Paginas: 57-61
Archivo PDF: 268.45 Kb.


RESUMEN

La proteína antiviral del ginseng (PAP), aislada de plantas de Phytolacca americana y Phytolacca acinosa, inhibe la traducción proteica mediante la remoción catalítica de un residuo de adenina específico, en la cadena mayor de la subunidad 60S del ARN ribosomal eucariótico. En este estudio se aisló y secuenció el gen PAP-I de P. americana, y posteriormente se comparó con los genes de otras proteínas inactivadoras de ribosomas (RIP), reportadas en GenBank®. Se extrajo el ADN total de las hojas tardías del verano de las plantas de P. americana y el fragmento de 868 pb correspondiente al ADN del gen se amplificó con el uso de cebadores específicos, mediante reacción en cadena de la polimerasa (PCR). El producto de la PCR eluido se purificó, se clonó en el vector pTZ57R/T, y se movilizó en células de Escherichia coli cepa DH5α. Tras la secuenciación del producto de la PCR del gen PAP-I, la secuencia mostró una homología nucleotídica de 98 a 82 % y aminoacídica de 94 a 26 %, con las RIP reportadas. El análisis filogenético confirmó que el gen amplificado corresponde a la RIP tipo I de simple cadena (PAP-I).


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