利用酵母GAL4双杂交系统分析Ras与CDC25鸟嘌呤核苷酸交换因子之间的相互作用。

Analysis of interaction between Ras and CDC25 guanine nucleotide exchange factor using yeast GAL4 two-hybrid system.

作者信息

Mosteller R D, Park W, Broek D

机构信息

Department of Biochemistry and Molecular Biology, Norris Comprehensive Cancer Center, University of Southern California School of Medicine, Los Angeles 90033, USA.

出版信息

Methods Enzymol. 1995;255:135-48. doi: 10.1016/s0076-6879(95)55017-8.

DOI:10.1016/s0076-6879(95)55017-8

PMID:8524097

Abstract

Our results demonstrate that the GAL4 two-hybrid system can be useful for studying interactions of the wild-type and mutant forms of Ras proteins with the CDC25 guanine nucleotide exchange factor (CDC25-GEF). In addition, our findings show that a negative result in the GAL4 two-hybrid system does not indicate that the two proteins tested do not interact under all conditions but only that they do not interact under the specific conditions examined. We recommend that the two-hybrid system be employed in combination with other approaches, including molecular genetic analyses and in vitro binding experiments, for the study of Ras and CDC25-GEF interactions.

摘要

我们的结果表明，GAL4双杂交系统可用于研究野生型和突变型Ras蛋白与CDC25鸟嘌呤核苷酸交换因子（CDC25-GEF）的相互作用。此外，我们的研究结果表明，GAL4双杂交系统中的阴性结果并不表明所测试的两种蛋白质在所有条件下都不相互作用，而仅表明它们在所检测的特定条件下不相互作用。我们建议将双杂交系统与其他方法结合使用，包括分子遗传学分析和体外结合实验，以研究Ras与CDC25-GEF的相互作用。