Production of dihydrofolate reductase by an improved continuous flow cell-free translation system using wheat germ extract.
We have examined the characteristics of protein synthesis in an improved continuous flow cell-free translation system prepared from wheat germ extract with dihydrofolate reductase mRNA as the translated message. Continuous buffer flow and separation of the product from the reaction mixture were accomplished by the use of a modified Amicon ultrafiltration chamber as the reaction vessel. The system worked for 19 hours and produced 1.52 nmol (27.4 micrograms) of enzymatically active dihydrofolate reductase.
Endo
Y
Y
Department of Biochemistry, Yamanashi Medical College, Japan.
Oka
T
T
Ogata
K
K
Natori
Y
Y
D001483:Base Sequence; D001709:Biotechnology; D002474:Cell-Free System; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis* / Q000235:genetics
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D013762:Tetrahydrofolate Dehydrogenase