Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China.
J Appl Microbiol. 2009 Mar;106(3):941-51. doi: 10.1111/j.1365-2672.2008.04063.x. Epub 2009 Jan 21.
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.
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.
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.
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.
研究酿酒酵母变体的青蒿基因编码的法呢基二磷酸合酶（ADS）对转化酵母中法呢烯-4,11-二烯产量的影响。
对 ADS 基因进行突变得到酵母变体 ADSm。通过一系列聚合酶链反应（PCR），逐步扩增基因的一小段区域来进行 ADSm 的合成。人工 ADSm 基因包含酵母翻译机制偏好的密码子。然后，该序列被整合到酵母表达载体 pYeDP60 中。融合构建体具有活性，与植物基因转化的酵母细胞相比，转化的酵母细胞产生了更高水平的法呢烯-4,11-二烯。
与转化植物基因的酵母相比，转化酵母变体（ADSm）的菌株在法呢烯-4,11-二烯的生产效率方面更具优势。
我们证明了通过串联 PCR 反应获得的外源基因的酵母变体可以解决酿酒酵母细胞中外源基因表达效率低的问题。
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