Expression Analysis In Vitro

Alfredo Colosimo, University of Rome ‘La Sapienza’, Rome, Italy

Published online: January 2006

DOI: 10.1038/npg.els.0005678

Expression analysis in vitro is a rapidly growing and constantly evolving field, based upon the use of a template of RNA, for a translation reaction, or of DNA, in a coupled transcription–translation system.

Keywords: reporter gene studies; DNase footprinting; methylation interference assays; gel-shift assay; yeast one-; two- and three-hybrid system; phage display

Figure 1. Schematic overview of yeast one-, two- and three-hybrid systems. DNA-BP and -AD are, respectively, the DNA-binding domain and the activation domain, identified in many eukaryotic transcriptional activators as functionally and physically independent units. (a) In one-hybrid systems, the two domains must be present in the same chimeric protein to allow generation of the transcriptional signal by the reporter gene, generally consisting in growth or color selection; (b) in two-hybrid systems, they are coupled to proteins P1 and P2, whose physical interaction is a necessary prerequisite for a successful transcription of the reporter gene; (c) in three-hybrid systems, a third hybrid molecule acts to bring together the DNA-BP fused to the receptor for one ligand with the DNA-AD fused to the receptor for the second ligand, thus reconstituting a functional transcriptional activator.
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Related Sub-Topics:

Protein Structure | Translation and Protein synthesis | Protein Folding, Evolution and Degradation | Replication and Recombination | Transcription | Transcription of DNA | Translation of RNA | Techniques and Tools in Molecular Biology | Enzymes: Structure and Action Mechanism | Nucleic Acids: Structure, Function, Metabolism
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Article Title: Expression Analysis In Vitro
 
How to Cite close
Colosimo, Alfredo(Jan 2006) Expression Analysis In Vitro. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0005678]