Synthesis of RNA having cap structure.
RNA consisting 43 nucleotides bearing cap structure was synthesized (Figure). In the first place, 9 mer of a leader sequence with the cap structure (F-1) was synthesized by the phosphotriester method and followed by the capping reaction. Next, 32 mer of a cistron was divided into two fragments and each was synthesized by the phosphoramidite method. The 3'-end nucleotide of the RNA, a modified guanosine 5'-phosphate, was introduced to F-3 by use of P1-2',3'-O-methoxymethylene guanosine-5'-yl P2-adenosine-5'-yl diphosphate (A5' ppGmM) with T4 RNA ligase. The chemically synthesized RNA fragments were ligated with T4 RNA ligase to afford the desired RNA.
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; D001483:Base Sequence; D008969:Molecular Sequence Data; D012313:RNA / Q000138:chemical synthesis*; D012315:RNA Caps / Q000737:chemistry*; D011130:RNA Ligase (ATP)
D012315:RNA Caps; D012313:RNA; D011130:RNA Ligase (ATP)