基于PCR方法敲除黄酒酵母精氨酸酶基因的工程菌构建

A PCR-based method for construction of a genetic engineering Chinese rice wine yeast by disrupting the CAR1 gene

摘要: 用一种高效快速的免克隆方法-长侧翼同源PCR（LFH-PCR）,构建基因敲除组件,然后通过化学转化方法把敲除组件转入黄酒酵母细胞中,利用同源重组机制精确敲除精氨酸酶基因（CAR1）,构建了黄酒酵母工程菌株。结果表明,敲除CAR1基因的工程菌与出发菌株相比,酒液中尿素的含量降低了72%,氨基甲酸乙酯含量降低了38%,并且该菌株的遗传稳定性好,发酵性能与出发菌株基本一致。

Abstract: Gene targeting cassettes were synthesized by long flanking homology PCR (LFH-PCR) , a new cloningfree strategy to delete genes, and were transformed into yeast.A engineering Chinese rice yeast was constructed by disruption of CAR1 gene.Compared to the parental wild type, the engineering strain exhibited reduced production of EC and urea in Chinese rice wine.They were reduced by 72%and 38%, respectively.It was also suggested that Chinese rice wine brewed with the recombinant yeast does not affect overall fermentation profiles.