Gene expression in a cell-free system on the preparative scale.
A cell-free system for preparative gene expression is described. It is composed of DNA-free Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20-50 h is hundreds of micrograms from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass.
Baranov
V I
VI
Insitute of Protein Research, Academy of Sciences of the USSR, Moscow Region.
Morozov IYu
Ortlepp
S A
SA
Spirin
A S
AS
D000255:Adenosine Triphosphate / Q000378:metabolism; D002021:Buffers; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D009711:Nucleotides; D010957:Plasmids; D014176:Protein Biosynthesis; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics
D002021:Buffers; D003470:Culture Media; D009711:Nucleotides; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D013762:Tetrahydrofolate Dehydrogenase; D001618:beta-Lactamases