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6660019 
Journal Article 
Chemical synthesis of a biologically active natural tRNA with its minor bases 
Gasparutto, D; Livache, T; Bazin, H; Duplaa, AM; Guy, A; Khorlin, A; Molko, D; Roget, A; Téoule, R; , 
1992 
1 
Nucleic Acids Research
ISSN: 0305-1048
EISSN: 1362-4962 
OXFORD UNIV PRESS 
OXFORD 
5159-5166 
English 
1383941 
WOS:A1992JV09200031 
The complete chemical synthesis of an E. coli tRNA(Ala) with its specific minor nucleosides, dihydrouridine, ribothymidine and pseudouridine, is reported. The method makes use of protected 2'-O-tertiobutyldimethylsilyl-ribonucleoside-3'-O-(2-cyanoethyl-N- ethyl-N- methyl)phosphoramidites. The exocyclic amino functions of the bases were protected by the phenoxyacetyl group for purines and acetyl for cytosine. The assembling has been performed on a silica support with coupling yield better than 98% within 2 min of condensation. Triethylamine tris-hydrofluoride allowed a clean and complete deprotection of the tBDMS groups. The synthetic tRNA(Ala) has been transcribed into cDNA by reverse transcriptase and sequenced. With E. coli alanyl-tRNA synthetase the alanyl acceptance activity and kcat/Km were 672 pmol/A260 and 6 x 10(4)M-1s-1, respectively.