High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier.
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.
Schiestl
R H
RH
Department of Biology, University of Rochester, NY 14627.
Gietz
R D
RD
D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D008722:Methods; D009691:Nucleic Acid Denaturation; D012331:RNA, Fungal / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013104:Spheroplasts / Q000378:metabolism; D013997:Time Factors; D014170:Transformation, Genetic
D004271:DNA, Fungal; D004277:DNA, Single-Stranded; D012331:RNA, Fungal