|
Download
article
150kb
pdf? |
|
The Next-Generation Assay for Mammalian Protein Interactions:
The CheckMate™/Flexi® Vector Mammalian Two-Hybrid System
Elaine Schenborn, Ph.D., Pete Stecha, B.S.,
Michael Slater, Ph.D., and Frank Fan, Ph.D.
Promega Corporation
Analyses of protein:protein interactions are an integral part of better
understanding protein functionality. Mammalian two-hybrid systems
provide an excellent means to detect and validate protein interactions
within mammalian cells. The next-generation CheckMate™/Flexi®
Vector Mammalian Two-Hybrid System provides additional features for
ease-of-use and expanded capabilities. The cloning vectors are now
compatible with the Flexi® Vector System, allowing easy
transfer of protein-coding regions to and from other Flexi®
Vectors, which contain specialized functionalities. The reporter vector
encodes firefly luciferase with rapid-response characteristics and a
hygromycin-resistance marker for generating stable cell lines. In this
article, we demonstrate the efficient transfer of protein-coding regions
into the CheckMate™/Flexi® Vectors and the ease of screening
for interacting partners. We also show how to use the mammalian
two-hybrid system to generate stable cell lines and modulate protein
interactions with small molecules in a stably transfected cell line.
|
