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Updated: Aug 26, 2026

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
Published on: February 7, 2019
In vitro folding and characterization of the p53 DNA binding domain
Christian Klein1, Friederike Hesse, Alexander Dehner
1Pharma Research, Nonnenwaldstrasse 2, Roche Diagnostics GmbH, D-82372 Penzberg, Germany. christian.klein.ck1@roche.com
Abstract:
The transcription factor p53 acts as major tumor suppressor and is inactivated by mutation in more than 50% of all human tumors. We have established an efficient procedure for the in vitro folding and purification of the p53 DNA binding domain (p53DBD) using a modified factorial matrix approach that supplies large amounts of homogeneous (isotope-labeled) p53DBD for application in biochemical, crystallographic and NMR spectroscopic studies. We further show with biophysical methods that in vitro folded p53DBD is fully functional and that its conformation is identical to that obtained from the soluble fraction.
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