Cell-free protein synthesis with prokaryotic combined transcription-translation.
Cell-free biology exploits and studies complex biological processes in a controlled environment without intact cells. One model system is prokaryotic cell-free protein synthesis. This technology offers an attractive and convenient approach to produce properly folded recombinant DNA (rDNA) proteins on a laboratory scale, screen PCR fragment libraries in a high-throughput format, express pharmaceutical proteins, incorporate labeled or unnatural amino acids into proteins, and activate microbial physiology to allow for investigation of biological systems. We describe the preparation of materials necessary for the expression, quantification, and purification of rDNA proteins from active Escherichia coli extracts.
Swartz
James R
JR
Department of Chemical Engineering, Stanford University, CA, USA.
Jewett
Michael C
MC
Woodrow
Kim A
KA
D001345:Autoradiography / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors; D010957:Plasmids; D014176:Protein Biosynthesis / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*; D014764:Viral Proteins
D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases