Schiestl R H, Gietz R D
Department of Biology, University of Rochester, NY 14627.
Curr Genet. 1989 Dec;16(5-6):339-46. doi: 10.1007/BF00340712.
A method, using LiAc to yield competent cells, is described that increased the efficiency of genetic transformation of intact cells of Saccharomyces cerevisiae to more than 1 X 10(5) transformants per microgram of vector DNA and to 1.5% transformants per viable cell. The use of single stranded, or heat denaturated double stranded, nucleic acids as carrier resulted in about a 100 fold higher frequency of transformation with plasmids containing the 2 microns origin of replication. Single stranded DNA seems to be responsible for the effect since M13 single stranded DNA, as well as RNA, was effective. Boiled carrier DNA did not yield any increased transformation efficiency using spheroplast formation to induce DNA uptake, indicating a difference in the mechanism of transformation with the two methods.
本文描述了一种使用醋酸锂制备感受态细胞的方法,该方法可将酿酒酵母完整细胞的遗传转化效率提高到每微克载体DNA产生超过1×10⁵个转化子,且每活细胞的转化子比例达到1.5%。使用单链或热变性双链核酸作为载体,可使含有2μm复制起点的质粒的转化频率提高约100倍。单链DNA似乎是产生这种效果的原因,因为M13单链DNA以及RNA都具有此效应。使用原生质球形成法诱导DNA摄取时,煮沸的载体DNA并未提高转化效率,这表明两种方法的转化机制存在差异。