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. 1992 Apr 11;20(7):1643–1648. doi: 10.1093/nar/20.7.1643

Molecular design of a eukaryotic messenger RNA and its chemical synthesis.

R Iwase 1, M Maeda 1, T Fujiwara 1, M Sekine 1, T Hata 1, K Miura 1
PMCID: PMC312250  PMID: 1374557

Abstract

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.

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Selected References

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