Cell-Free And Happy: In Vitro Translation And Transcription/Translation Systems

Date: June 21, 1999Kits for In Vitro Translation and Transcription/Translation Systems Problem proteins--you know the ones. Some are rapidly degraded by endogenous proteins. Others are toxic to the very cells used to overexpress them. These types of problems can sometimes be avoided by using cell-free extracts for the in vitro expression of proteins. In vitro translation systems are also useful for other applications, including incorporating modified or unnatural amino acids for functional st

Deborah Wilkinson
Jun 20, 1999

Date: June 21, 1999Kits for In Vitro Translation and Transcription/Translation Systems

Problem proteins--you know the ones. Some are rapidly degraded by endogenous proteins. Others are toxic to the very cells used to overexpress them. These types of problems can sometimes be avoided by using cell-free extracts for the in vitro expression of proteins. In vitro translation systems are also useful for other applications, including incorporating modified or unnatural amino acids for functional studies and localizing mutations via synthesis of truncated gene products.

In "conventional" systems, RNA is used directly as a template for translation. The RNA can be total RNA, mRNA, or a synthesized template. "Linked" or "coupled" translation systems start off with a DNA template. RNA is transcribed from the DNA template, then subsequently translated without purification. Appropriate synthetic DNA templates for in vitro transcription/translation systems are readily generated, either after cloning into plasmid vectors or via PCR....

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