Open AccessTotal synthesis of a RNA molecule with sequence identical to that of Escherichia coli formylmethionine tRNA.
Author(s) -
Eiko Ohtsuka,
Shoji Tanaka,
Toshiki Tanaka,
Toru Miyake,
Alexander F. Markham,
Eiko Nakagawa,
Toshiaki Wakabayashi,
Yoshio Taniyama,
Satoshi Nishikawa,
Ryoichi Fukumoto,
Haruki Uemura,
Takefumi Doi,
Toko Tokunaga,
Morio Ikehara
Publication year - 1981
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.78.9.5493
Subject(s) - rna ligase , phosphodiester bond , oligonucleotide , transfer rna , escherichia coli , polynucleotide , rna , aminoacylation , biochemistry , dna ligase , chemistry , molecule , biology , stereochemistry , enzyme , dna , gene , organic chemistry
A RNA molecule has been synthesized that is identical in sequence to Escherichia coli tRNAfMet except that it lacks the base modifications present in the E. coli tRNA. This was achieved by enzymatic joining of chemically synthesized oligonucleotides with chain lengths of 3-10 which were synthesized by the phosphodiester or phosphotriester method. First, quarter molecules of tRNA were constructed by joining of chemically synthesized fragments with RNA ligase. The 5'-quarter molecule (bases 1-20) served as an acceptor in joining reactions with the 3',5'-bisphosphorylated donor molecule (bases 21-34). The 5'-half molecule thus obtained was treated with phosphatase and joined to the 3'-half molecule which was prepared by ligation of the other quarter molecules (bases 35-60, acceptor; bases 61-77, donor) followed by 5'-phosphorylation with polynucleotide kinase. The synthetic tRNA was characterized by oligonucleotide pattern and was partially active in aminoacylation with E. coli methionyl-tRNA synthetase.