HIGH-EFFICIENCY TRANSFORMATION OF INTACT YEAST-CELLS USING SINGLE STRANDED NUCLEIC-ACIDS AS A CARRIER
HIGH-EFFICIENCY TRANSFORMATION OF INTACT YEAST-CELLS USING SINGLE STRANDED NUCLEIC-ACIDS AS A CARRIER
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DOI：
10.1007/bf00340712
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发表时间：
1989-12-01
期刊：
影响因子：
2.5
通讯作者：
GIETZ, RD
中科院分区：
文献类型：
--
作者：
SCHIESTL, RH;GIETZ, RD
本文描述了一种利用醋酸锂（LiAc）制备感受态细胞的方法，该方法可将酿酒酵母（Saccharomyces cerevisae）完整细胞的遗传转化效率提高到每微克载体DNA超过1×10⁵个转化子，以及每活细胞1.5%的转化子。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都有效。使用原生质体形成来诱导DNA摄取时，煮沸的载体DNA并没有提高转化效率，这表明两种转化方法的机制存在差异。
A method, using LiAc to yield competent cells, is described that increased the efficiency of genetic transformation of intact cells of Saccharomyces cerevisae to more than 1 .times. 105 transformants per microgam 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 .mu.m 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.