Production of amorpha-4,11-diene in engineered yeasts / 药学学报
Acta Pharmaceutica Sinica
;
(12): 1297-1303, 2009.
Artículo
en Chino
| WPRIM
| ID: wpr-344082
ABSTRACT
Plasmid-carrying Saccharomyces cerevisia (W303-1B[pYeDP60/G/ADS]) and genome-transformed S. cerevisia (W303-1B[rDNAADS]), both harboring amorpha-4,11-diene synthase (ADS) gene were constructed to investigate the production of amorpha-4,11-diene. The recombinant plasmid pYeDP60/G/ADS that harbors the ADS gene was transformed into S. cerevisiae W303-1B, resulting in the engineered yeast W303-1B[pYeDP60/G/ADS], which contains multi-copies of the plasmid. The ADS gene expression cassette was obtained by PCR amplification of the pYeDP60/G/ADS template, and then introduced into S. cerevisiae W303-1B to obtain the engineered yeast W303-1B[rDNAADS], in which the ADS gene was integrated into the rDNA locus of the yeast genome through the homologous recombination. GC-MS analysis confirmed that both of the engineered yeasts could produce amorpha-4,11-diene. Moreover, the amorpha-4,11-diene yield of W303-1B[pYeDP60/G/ADS] was higher than that of W303-1B[rDNAADS]. Southern blot analysis showed that there is only one copy of ADS gene in the genome of W303-1B[rDNAADS]. It implied that the amorpha-4,11-diene yield can be improved by increasing the ADS gene copies.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
Saccharomyces cerevisiae
/
Sesquiterpenos
/
Transformación Genética
/
ADN Ribosómico
/
Ingeniería Genética
/
Genoma Fúngico
/
Transferasas Alquil y Aril
/
Fermentación
/
Genética
Idioma:
Chino
Revista:
Acta Pharmaceutica Sinica
Año:
2009
Tipo del documento:
Artículo
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