Synthesis of 5′ Fragments of Formylmethionine Transfer Ribonucleic Acid and Their Reconstitution with a Natural Three‐Quarter Molecule

Publisher Website
Google Scholar
Abstract
An eicosanucleotide C-G-C-G-G-G-G-U-G-G-A-G-C-A-G-C-C-U-G-Gp corresponding to the bases 1-20 of the nascent sequence for the Escherichia coli tRNAfMet was synthesized by the joining of the chemically synthesized oligonucleotides C-G-C-G, G-G-G-U-G-G and A-G-C-A-G-C-C-U-G-Gp using RNA ligase from [phage] T4-infected E. coli. The hexanucleotide and decanucleotide were phosphorylated with polynucleotide kinase and [.gamma.-32P]ATP prior to the joining reactions. The decanucleotide and eicosanucleotide were reconstituted, respectively, with the 3''-three-quarter molecule obtained by limited digestion with RNase T1 of the natural tRNAfMet from E. coli and the activity of the complex as a methionine acceptor was tested using purified methionyl-tRNA synthetase from E. coli. The amino acid acceptor activity of the reconstituted molecules was 11% and 84% with respect to that of the intact tRNAfMet.

This publication has 21 references indexed in Scilit: