Reconstruction of tRNA^Phe molecules from the fragments by linkage with T‐4 RNA ligase in double‐stranded regions

Abstract

Phenylalanine-specific tRNA from yeast was hydrolysed with cobra venom ribonuclease in the doublestranded regions and the fragments isolated. The 'dissected' molecules with nicks in positions 28 and 41were reconstructed from supplementary fragments and treated with T-4 RNA ligase. A phosphodiester bond between two fragments was formed when the fragment combination (l-28) + (29-76) was used. A strong discrimination in the ligation yield between different nick positions in the same helix is shown.Cobra venom nuclease RNA ligase