Total synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.

• DOI: 10.1073/pnas.78.9.5493
• Publication Date: 2006-05-31T08:49:43Z
• Authors: E Ohtsuka; S Tanaka; T Tanaka; T Miyake; A F Markham; E Nakagawa; T Wakabayashi; Y Taniyama; S Nishikawa; R Fukumoto; H Uemura; T Doi; T Tokunaga; M Ikehara
• Keywords: RNA ligase; Phosphodiester bond; Polynucleotide; Aminoacylation

A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNAfMet except it lacks the base modifications present E. tRNA. This was achieved by enzymatic joining of chemically oligonucleotides with chain lengths 3-10 which were phosphodiester or phosphotriester method. First, quarter molecules tRNA constructed fragments ligase. The 5'-quarter (bases 1-20) served as an acceptor reactions 3',5'-bisphosphorylated donor 21-34). 5'-half thus obtained treated phosphatase and joined 3'-half prepared ligation other 35-60, acceptor; bases 61-77, donor) followed 5'-phosphorylation polynucleotide kinase. synthetic characterized oligonucleotide pattern partially active aminoacylation methionyl-tRNA synthetase.