1 research outputs found
ARTICULO ORIGINAL CORTO / SHORT ORIGINAL PAPER - HIGH LEVEL EXPRESSION OF HUMAN IFN-alpha2b IN Pichia Pastoris
The gene coding for human interferon-alpha2b was integrated into the Pichia pastoris genome. The expression of the interferon gene was controlled by methanol-regulated alcohol oxidase (AOX1) promoter. An expression level of 400 mg of interferon per liter of culture was achieved under appropriate fermentation conditions. The molecule was purified to homogeneity and partially characterized in terms of biological activity and structural properties. Fast Atom Bombardment Mass Spectrometry (FAB-MS) allowed the verification of the aminoacid sequence and indicated the presence of amino-terminal acetyl groups in 70% of the final product. An alternative to eliminate this modification is suggested.
RESUMEN
El gen que codifica para el interferon-alfa 2b humano fue integrado en el genoma de Pichia pastoris. La expresion del gen de interferon fue controlada por la induccion con metanol del promotor alcohol oxidasa 1. Se alcanzaron 400 mg de interferon por litro de cultivo en fermentacion. La molecula fue purificada y parcialmente caracterizada. Se verifico la secuencia aminoacidica por espectrometria de masas con ionizacion por bombardeo con atomos acelerados (FAB-MS) detectandose un 70% de acetilacion. Una alternativa para eliminar esta modificacion es propuesta