Molecular design of a eukaryotic messenger RNA and its chemical synthesis.
A designed mRNA consisting of 42 ribonucleotides having the cap structure was synthesized. The capped leader sequence of the brome mosaic virus (BMV) mRNA 4, m7G5'pppGUAUUAAUA (F-1), was synthesized by the phosphotriester method and followed by the capping reaction. A 32-mer consisting of an initiation codon (AUG), the coding region corresponding to a bacterial pheromone cAD1 and two stop codons, was constructed by the 18-mer (F-2) and 14-mer (F-3), which were synthesized by the phosphoramidite method. 2'-,3'-O-Methoxymethylene-guanosine 5'-phosphate was condensed with F-3 using P1-2',3'-O-methoxymethyleneguanosine-5'-yl P2-adenosine-5'-yl pyrophosphate (9) with T4 RNA ligase. The chemically synthesized RNA fragments were ligated successively with T4 RNa ligase to afford the whole RNA molecule.
Iwase
R
R
Department of Life Chemistry, Tokyo Institute of Technology, Yokohama, Japan.
Maeda
M
M
Fujiwara
T
T
Sekine
M
M
Hata
T
T
Miura
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics; D006151:Guanosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics; D009842:Oligopeptides / Q000235:genetics; D010675:Pheromones / Q000235:genetics; D010766:Phosphorylation; D012315:RNA Caps / Q000138:chemical synthesis* / Q000235:genetics; D011130:RNA Ligase (ATP) / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000138:chemical synthesis* / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics
D001426:Bacterial Proteins; D003062:Codon; D009842:Oligopeptides; D010675:Pheromones; D012315:RNA Caps; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012367:RNA, Viral; D006151:Guanosine; C016578:7-methylguanosine; C043691:cAD1 bacterial sex hormone; D011130:RNA Ligase (ATP)