Schneider B L, Steiner B, Seufert W, Futcher A B
Cold Spring Harbor Laboratory, NY 11724, USA.
Yeast. 1996 Feb;12(2):129-34. doi: 10.1002/(sici)1097-0061(199602)12:2<129::aid-yea891>3.0.co;2-o.
Gene disruption is an important method for genetic analysis in Saccharomyces cerevisiae. We have designed a polymerase chain reaction-directed gene disruption cassette that allows rapid disruption of genes in S. cerevisiae without previously cloning them. In addition, this cassette allows recycling of URA3, generating gene disruptions without the permanent loss of the ura3 marker. An indefinite number of disruptions can therefore be made in the same strain.
基因破坏是酿酒酵母遗传分析的一种重要方法。我们设计了一种聚合酶链反应导向的基因破坏盒,它能够在不预先克隆基因的情况下快速破坏酿酒酵母中的基因。此外,这个盒允许URA3循环利用,从而在不永久丢失ura3标记的情况下产生基因破坏。因此,可以在同一菌株中进行无限次的破坏。