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The improvement of amorpha-4,11-diene production by a yeast-conform variant

2009; Oxford University Press; Volume: 106; Issue: 3 Linguagem: Inglês

10.1111/j.1365-2672.2008.04063.x

1365-2672

K. Jiang-Qiang, Wei Wang, W. Li-Na, Zhang Xiaodong, Cheng Kedi, Ping Zhu,

Fungal and yeast genetics research

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.