Rapid translation system: a novel cell-free way from gene to protein.
Proteome research has recently been stimulated by important technological advances in the field of recombinant protein expression. One major breakthrough was the development of a new generation of cell-free transcription/translation systems. The open and flexible character of these systems allows direct control over expression conditions via the addition of supplements to the expression reaction. The possibility of working with linear expression templates instead of cloned plasmids and the ease of downstream processing, circumventing the need for cell-lysis, makes them ideally suited for high-throughput screening applications. Among these novel cell-free systems, the Rapid Translation System (RTS) developed by Roche is the first one that is scalable from micrograms to milligrams of protein. This review describes the basic principles of RTS which differentiate it from traditional in vitro expression technologies, starting from template generation to high-end applications like labeling for structural biology research. Recent results obtained by RTS users from different institutions are presented to illustrate each step of a novel cell-free protein expression workflow and its benefits compared to traditional cell-based expression.
Hoffmann
Michael
M
Roche Diagnostics GmbH, Nonnenwald 2, 82372 Penzberg, Germany. michael.hoffmann@roche.com
Nemetz
Cordula
C
Madin
Kairat
K
Buchberger
Bernd
B
D002457:Cell Extracts; D002474:Cell-Free System; D004926:Escherichia coli; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D014176:Protein Biosynthesis; D017510:Protein Folding; D046188:Protein Modification, Translational / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors; D014158:Transcription, Genetic
D002457:Cell Extracts; D011994:Recombinant Proteins