Construction of an efficient expression vector for coupled transcription/translation in a wheat germ cell-free system.
Using the expression vector, pEU, which we have constructed, highly efficient in vitro protein synthesis can be achieved: The system works for 150 hours and without further template addition once the reaction has started, yielding 5 mg of enzymatically active protein in a 1 ml reaction.
Sawasaki
T
T
Department of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama 790-8577, Japan.
Hasegawa
Y
Y
Tsuchimochi
M
M
Kasahara
Y
Y
Endo
Y
Y
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D001483:Base Sequence; D001709:Biotechnology; D002474:Cell-Free System; D005822:Genetic Vectors; D007700:Kinetics; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic; D014908:Triticum / Q000235:genetics* / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D012314:RNA Cap Analogs; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014764:Viral Proteins; D013762:Tetrahydrofolate Dehydrogenase