Production of an enzymatic active protein using a continuous flow cell-free translation system.
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 (dhfr) mRNA as the translated message. Continuous buffer flow and separation of product from the reaction mixture were accomplished by the use of a modified Amicon ultrafiltration chamber as reaction vessel. The system produced protein for more than 20 h, and the product had an activity of dhfr comparable to that of authentic enzyme from E. coli. Analysis of RNA recovered from the filtrate supports the notion that a functionally active protein-synthesizing machinery is superorganized in a dynamic complex.
Department of Biochemistry, Yamanashi Medical College, Japan.
Y
Y
Endo
S
S
Otsuzuki
K
K
Ito
K
K
Miura
D001483:Base Sequence; D002474:Cell-Free System; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
D012333:RNA, Messenger; D013762:Tetrahydrofolate Dehydrogenase