Applications of the TNT® T7 Quick
Coupled Transcription/Translation System
Gregory S. Beckler
Promega Corporation
Use of the TNT® T7 Quick System for the Detection of
Translation-Terminating Mutations in a Protein Truncation Test
Matthew Cayouette
LabCorp of America
1912 Alexander Drive
Research Triangle Park, NC 27709
Protein truncation tests are powerful research methods for
detecting frameshift and nonsense mutations in many genes. In this update, we describe the
use of the TNT®
T7 Quick System(a) to characterize proteins produced by normal and
mutant forms of the APC gene, which is responsible for familial adenomatous
polyposis (FAP).
Analysis of Interactions between Proteinases and Serpins Expressed in vitro Using
the TNT® T7 Quick System
Peter C. Turner
Department of Molecular Genetics and Microbiology
University of Florida
Gainesville, FL 32610
The interactions between proteinases and natural inhibitory
proteins (serpins) can be conveniently studied using radiolabeled serpins expressed
in
vitro with the TNT®
T7 Quick Coupled Transcription/Translation System(a). Most serpins can be
efficiently expressed using the standard TNT® T7 Quick System protocol. However, some serpins, and
presumably other proteins, are not expressed efficiently unless additional magnesium is
included in the coupled transcription/translation reaction. We also demonstrate that
serpins synthesized using the TNT® T7 Quick System are functionally active in protein-protein
interactions.
|
