Lue N F, Chasman D I, Buchman A R, Kornberg R D
Department of Cell Biology, Stanford University School of Medicine, California 94305.
Mol Cell Biol. 1987 Oct;7(10):3446-51. doi: 10.1128/mcb.7.10.3446-3451.1987.
The GAL80 protein of Saccharomyces cerevisiae, synthesized in vitro, bound tightly to GAL4 protein and to a GAL4 protein-upstream activation sequence DNA complex, as shown by (i) coimmunoprecipitation of GAL4 and GAL80 proteins with anti-GAL4 antiserum, (ii) an electrophoretic mobility shift of a GAL4 protein-upstream activation sequence DNA complex upon the addition of GAL80 protein, and (iii) GAL4-dependent binding of GAL80 protein to upstream activation sequence DNA immobilized on Sepharose beads. Anti-GAL4 antisera were raised against a GAL4-URA3 fusion protein, which could be purified to homogeneity in a single step with the use of an affinity chromatographic procedure for the URA3 gene product.
体外合成的酿酒酵母GAL80蛋白与GAL4蛋白以及GAL4蛋白-上游激活序列DNA复合物紧密结合，这通过以下方式得以证明：（i）用抗GAL4抗血清对GAL4和GAL80蛋白进行共免疫沉淀；（ii）加入GAL80蛋白后GAL4蛋白-上游激活序列DNA复合物的电泳迁移率发生改变；（iii）GAL80蛋白与固定在琼脂糖珠上的上游激活序列DNA的GAL4依赖性结合。抗GAL4抗血清是针对GAL4-URA3融合蛋白产生的，该融合蛋白可通过使用针对URA3基因产物的亲和色谱程序一步纯化至同质。
