Unit

UNIT 10.17 Synthesizing Proteins In Vitro by Transcription and Translation of Cloned Genes

  1. Kevin Struhl

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1017s48

Current Protocols in Molecular Biology

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.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1999

Abstract

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.