Smith V, Botstein D, Brown P O
Department of Genetics, Stanford University, CA, USA.
Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6479-83. doi: 10.1073/pnas.92.14.6479.
This report describes an efficient strategy for determining the functions of sequenced genes in microorganisms. A large population of cells is subjected to insertional mutagenesis. The mutagenized population is then divided into representative samples, each of which is subjected to a different selection. DNA is prepared from each sample population after the selection. The polymerase chain reaction is then used to determine retrospectively whether insertions into a particular sequence affected the outcome of any selection. The method is efficient because the insertional mutagenesis and each selection need only to be performed once to enable the functions of thousands of genes to be investigated, rather than once for each gene. We tested this "genetic footprinting" strategy using the model organism Saccharomyces cerevisiae.
本报告描述了一种用于确定微生物中已测序基因功能的有效策略。大量细胞群体接受插入诱变。诱变后的群体随后被分成代表性样本，每个样本接受不同的筛选。筛选后从每个样本群体中制备DNA。然后使用聚合酶链反应来追溯确定插入特定序列是否影响任何筛选的结果。该方法很有效，因为插入诱变和每次筛选只需要进行一次，就能研究数千个基因的功能，而不必对每个基因都进行一次。我们使用模式生物酿酒酵母测试了这种“遗传足迹”策略。
