Open AccessRNA-Linked DNA Fragments In Vitro
Author(s) -
Akio Sugino,
Reiji Okazaki
Publication year - 1973
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.70.1.88
Subject(s) - rna , dna , rnase p , chemistry , rnase h , microbiology and biotechnology , nuclease protection assay , ribozyme , escherichia coli , nucleoside , biochemistry , biology , non coding rna , gene
RNA-linked DNA fragments are intermediates in DNA replication inEscherichia coli cells made permeable to nucleoside triphosphates by treatment with toluene. Covalent linkage of a short RNA stretch to the 5′ end of the DNA is proved by transfer of32 P from [α-32 P]dNTP to ribonucleotides upon digestion with alkali or pancreatic RNase, and by a small decrease in the molecular size upon alkaline hydrolysis. The32 P transfer experiments reveal a unique structure...p(rPy)p(rA)p(rU or rC)p(dC)p... at the RNA-DNA junction.