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J Appl Microbiol ; 106(3): 941-51, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19191957

ABSTRACT

AIMS: To investigate the effect of the yeast-conform variant of the Artemisia annua gene encoding for amorpha-4,11-diene synthase (ADS) on the production of amorpha-4,11-diene in a transformed yeast. METHODS AND RESULTS: The ADS gene was mutated to the yeast-conform variant ADSm. The ADSm synthesis was performed based on step-by-step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artificial ADSm gene contained codons preferred by the yeast translation machinery. The sequence was then integrated into a yeast expression vector pYeDP60. The fusion construct was active and the transformed yeast cells produced higher level of amorpha-4,11-diene compared with the plant gene-transformed yeast cells. CONCLUSIONS: Strains transformed with the yeast-conform allele (ADSm) were more efficient in terms of production of amorpha-4,11-diene than those transformed with the plant gene. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrated that yeast-conform allele of foreign genes by serial PCR reactions can be a solution to low efficiency of heterologous gene expression in Saccharomyces cerevisiae cells.


Subject(s)
Alkyl and Aryl Transferases/metabolism , Artemisia annua/enzymology , Genetic Engineering/methods , Saccharomyces cerevisiae/genetics , Sesquiterpenes/metabolism , Alkyl and Aryl Transferases/genetics , Artemisia annua/genetics , Artemisia annua/metabolism , Gas Chromatography-Mass Spectrometry , Genome, Fungal/genetics , Molecular Sequence Data , Plasmids/genetics , Polycyclic Sesquiterpenes , Polymerase Chain Reaction , Recombination, Genetic , Saccharomyces cerevisiae/metabolism , Transformation, Genetic
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