UNIT 10.17 Synthesizing Proteins In Vitro by Transcription and Translation of Cloned Genes
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Struhl, K. 2001. Synthesizing Proteins In Vitro by Transcription and Translation of Cloned Genes. Current Protocols in Molecular Biology. 48:VI:10.17:10.17.1–10.17.5.
- Published Online: 1 MAY 2001
- Published Print: OCT 1999
The availability of a cloned gene makes it possible to synthesize the encoded protein by in vitro transcription and translation. The basis of the method presented in this unit is to clone the protein-coding sequences into a vector containing a promoter for SP6 or T7 RNA polymerase, to produce messenger RNA by transcribing the DNA template with this enzyme, and to synthesize the desired protein (often as an 35S-labeled species) by translation of this mRNA in vitro. A major advantage of this method is that any desired mutant protein can be generated simply by altering the DNA template.