知识与财富的链接
构建一株酿酒酵母SNF4基因缺失菌株并研究其对乙醇产量的影响。扩增带有SNF4基因上下游同源序列和Kanr筛选标记的SNF4基因敲除组件,转化到酿酒酵母YS2获得阳性克隆子,然后将质粒pSH65转到阳性克隆子中,半乳糖诱导pSH65表达Cre酶切除Kanr筛选标记,获得SNF4等位基因完全缺失菌株YS2-△SNF4。发酵实验结果表明,缺失菌株YS2-△SNF4乙醇产量较出发菌株提高了7.57％。利用Cre-LoxP系统,成功构建了SNF4等位基因完全缺失菌株并提高乙醇产生量。
Construction and ethanol production effects of SNF4 gene knockout in Saccharomyces cerevisiae were described in this paper.For knockout of SNF4 gene in S.cerevisiae YS2,a PCR-amplified disruption cassette was used,encoding the short flanking homologous regions to the SNF4 gene and Kanr as selectable marker.The SNF4 gene disruption cassette was transformed into S.cerevisiae YS2 through LiAc/SS Carrier DNA/PEG.The positive transformants were grown on G418 plates and verified by PCR.The Kanr marker was rescued...
