Production of amorpha-4,11-diene in engineered yeasts / 药学学报
Yao Xue Xue Bao
; (12): 1297-1303, 2009.
Article
de Zh
| WPRIM
| ID: wpr-344082
Bibliothèque responsable:
WPRO
ABSTRACT
Plasmid-carrying Saccharomyces cerevisia (W303-1B[pYeDP60/G/ADS]) and genome-transformed S. cerevisia (W303-1B[rDNA:ADS]), 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[rDNA:ADS], 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[rDNA:ADS]. Southern blot analysis showed that there is only one copy of ADS gene in the genome of W303-1B[rDNA:ADS]. It implied that the amorpha-4,11-diene yield can be improved by increasing the ADS gene copies.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Plasmides
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Saccharomyces cerevisiae
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Sesquiterpènes
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Transformation génétique
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ADN ribosomique
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Génie génétique
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Génome fongique
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Alkyl et aryl transferases
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Fermentation
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Génétique
langue:
Zh
Texte intégral:
Yao Xue Xue Bao
Année:
2009
Type:
Article