菌落PCR快速扩增工业酿酒酵母基因组DNA片段

Rapid Amplification of Genome DNA Fragment of Industrial Saccharomyces cerevisiae by Colony PCR

针对工业酿酒酵母细胞破壁难和提取基因酵母组DNA费时长(2-3 h/样品)的问题,研究了SDS-微珠涡旋破壁的方法。以破壁液上清为模板进行菌落PCR扩增酿酒酵母南阳K基因组中铜抗性蛋白基因(cup1)片段和rDNA片段。结果表明,在不需要提取酵母基因组前提下,利用该方法能有效地扩增酵母基因组DNA片段,同时该方法也适用于对转基因酿酒酵母进行菌落PCR从而实现对转化子的准确和快速地鉴定筛选,是一种成本低和易于操作的适于处理大量样品的方法。

For the difficulty of disrupting the cell of industrial Saccharomyces cerevisiae and time-consuming（2-3 h/sample） to extract yeast genome,we studied SDS-mini glass beads vortex method to disrupt the cell of industrial Saccharomyces cerevisiae.Supernatant of cell lysates was used as templet for colony PCR to amplify copper-resistant gene CUP1 fragment and ribosomal DNA（rDNA） fragment of Saccharomyces cerevisiae.The results showed that yeast genome DNA fragments could be amplified effectively without extraction of yeast genome.The method which was proved easy and cheap as performed also could be used for colony PCR to screen the transformants of genetically modified yeast accurately and rapidly.