一株酿酒酵母的特征区域序列扩增标记

以23株不同来源的酿酒酵母为研究对象,分别提取基因组DNA,试用50条随机引物对其进行随机扩增多态性DNA分析,筛选到两条具有菌株鉴别能力的随机引物。P09可以从2.412,ST—01,SK—26,ZD—01四株酿酒酵母基因组DNA中扩增出长度为433bp的Sc—433片断Sc—433;P46可以从2.1882,ST—01,NJ—02三株酿酒酵母基因组DNA中扩增出长度为665bp的Sc—665片断,其中仅有目的菌株ST—01能稳定的扩增出这2个标记。把这2个片断分别克隆到pUCmT质粒载体中,经过酶切鉴定后测序,根据序列设计特异性引物,把RAPD标记转化为特征区域序列扩增标记,为酿酒酵母菌株的分子鉴别提供了新的借鉴。

Identification of one Saccharomyces cerevisiae Strain by Sequenced Characterized Amplified Region Markers

In this paper,50 random primers were tested to the random amplified polymorphic DNA (RAPD)analysis on 23 strains of Saccharomyces cerevisiae isolated from different sources,and 2 effective primers named P09 and P46 were selected. Among the tested strains,4 strains(2.412,ST-01,SK-26,ZD- 01),have Sc-433 of 433bp fragment,were amplified with P09,and 3 strains(2.1882,ST-01,NJ-02),have Sc-665 of 665bp fragment,were amplified with P46,only the strain of ST-01 has the two markers simulta- neity.These two amplified DNA fragments were cloned into pUCmT plasmid and sequenced after identifica- tion of double digestion.Two pairs of specific primers were designed according to the sequence of amplified DNA fragments,and the sequenced characterized amplified region(SCAR)markers of ST—01 were devel- oped.These results could afford some new idea for molecular markers of Saccharomyces cerevisiae.