基础医学模式生物酿酒酵母接合型的鉴定——PCR法

Identification of the mating-type of model organism S. cerevisiae in basic medical science by PCR

目的:利用PCR法鉴定酿酒酵母a、α单倍体或a/α双倍体三种接合型。方法:根据酿酒酵母a、α或a/α三种接合型MAT基因座及附近序列信息,分别设计特异的寡核苷酸引物。提取酿酒酵母DNA作为模版和利用单菌落PCR法,进行特异性PCR扩增。结果:MATa基因座特异性引物能在a和a/α接合型中扩增出特异性条带;MATα基因座特异性引物则能在α和a/α接合型中扩增出特异性条带。结论:利用PCR鉴定酿酒酵母接合型是一种操作简单、准确灵敏的方法,为基础医学单细胞真核模式生物的研究提供方便。

Objective： To identify S. cerevisiae mating-type to be a, α haploid or a/α diploid by PCR. Methods： Specific PCR primers were designed according to the MAT locus and its nearby sequence of the three mating-types, and applied in specific PCR reaction using genome DNA as template or the method of colony PCR. Results： A specific band was obtained in a or a/α mating-type when specific primers were used on MATa, while a specific band was obtained in α or a/α mating-type by using specific primers on MATα. Conclusion： With specific primers, PCR may be a convenient, fast and precise method to identify S. cerevisiae mating-type, which could facilitate the study of monocellular eukaryotes model organism in basic medical science.