Blanc H, Dujon B, Guerineau M, Slonimski P P
Mol Gen Genet. 1978 May 31;161(3):311-5. doi: 10.1007/BF00331006.
A procedure is described for the detection of specific DNA sequences in Saccharomyces cerevisiae. This method allows a rapid screening of a large number of yeast colonies. The yeast cells of each colony, grown on nitrocellulose filters, are converted, in situ, to protoplasts by snail enzyme, and are then lysed and their DNAs are denatured and fixed on the filter. The presence of the specific DNA sequence is detected directly on the filter by hybridization with a radioactive cRNA. We have used successfully this technique to detect the presence or the absence of specific mt DNA sequences in p+, p- and p0 strains, and to detect the presence or the absence of the 2 mum DNA sequences in different strains.
本文描述了一种用于检测酿酒酵母中特定DNA序列的方法。该方法可快速筛选大量酵母菌落。在硝酸纤维素滤膜上生长的每个菌落的酵母细胞，通过蜗牛酶原位转化为原生质体，然后裂解，其DNA变性并固定在滤膜上。通过与放射性cRNA杂交直接在滤膜上检测特定DNA序列的存在。我们已成功使用该技术检测p+、p-和p0菌株中特定线粒体DNA序列的有无，以及不同菌株中2μm DNA序列的有无。
