Reconstruction of tRNAPhe molecules from the fragments by linkage with T-4 RNA ligase in double-stranded regions.
Phenylalanine-specific tRNA from yeast was hydrolysed with cobra venom ribonuclease in the double-stranded regions and the fragments isolated. The 'dissected' molecules with nicks in positions 28 and 41 were reconstructed from supplementary fragments and treated with T-4 RNA ligase. A phosphodiester bond between two fragments was formed when the fragment combination (1-28) + (29-76) was used. A strong discrimination in the ligation yield between different nick positions in the same helix is shown.
D001483:Base Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D006868:Hydrolysis; D011118:Polynucleotide Ligases / Q000378:metabolism*; D011130:RNA Ligase (ATP) / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic; D012441:Saccharomyces cerevisiae / Q000032:analysis; D013604:T-Phages / Q000201:enzymology*
D012346:RNA, Transfer, Amino Acyl; D012259:Ribonuclease, Pancreatic; D011118:Polynucleotide Ligases; D011130:RNA Ligase (ATP)