Cell-free translation system using phosphorothioate-containing mRNA.
Phosphorothioate-containing RNAs were generated by transcription of template DNA using the Sp diastereomers of ribonucleoside 5'-O-(1-thiotriphosphates) (NTP alpha S) and T7 RNA polymerase. The substitution of mRNA by phosphorothioate increased the efficiency of protein synthesis by stabilizing the mRNAs in prokaryotic cell-free translation systems. The substituted mRNAs were also shown to be applicable to the continuous cell-free translation system developed by Spirin and coworkers.
Ueda
T
T
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Tohda
H
H
Chikazumi
N
N
Eckstein
F
F
Watanabe
K
K
D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D016963:Thermus thermophilus / Q000235:genetics; D013873:Thionucleotides / Q000737:chemistry*
D012329:RNA, Bacterial; D012333:RNA, Messenger; D013873:Thionucleotides; D013762:Tetrahydrofolate Dehydrogenase