The improvement of amorpha-4,11-diene production by a yeast-conform variant |
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Authors: | K Jiang-Qiang W Wei W Li-Na Z Xiao-Dong C Ke-Di Z Ping |
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Institution: | Institute of Materia Medica, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, P.R. China |
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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. |
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Keywords: | amorpha-4 11-diene synthase Artemisia annua artemisinin Saccharomyces cerevisiae yeast-conform variant |
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