DNA Nucleotide Sequence Restricted by the RI Endonuclease

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 69; no. 11; pp. 3448 - 3452
• Main Authors: Hedgpeth, Joe, Goodman, Howard M., Boyer, Herbert W.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01-11-1972; National Acad Sciences
• Subjects: Adenine Nucleotides - analysis; Alkaline Phosphatase - metabolism; Avian Sarcoma Viruses - enzymology; Base Sequence; Biochemistry; Biological Sciences: Biochemistry; Coliphages; Cytosine Nucleotides - analysis; Deoxyribonucleases - metabolism; DNA; DNA Nucleotidyltransferases - metabolism; DNA, Viral - biosynthesis; DNA, Viral - metabolism; Drug Resistance, Microbial; Electrophoresis; Endonucleases - metabolism; Enzymes; Escherichia coli - enzymology; Extrachromosomal Inheritance; Genomes; Molecules; Nucleotide sequences; Nucleotides; Nucleotides - analysis; Nucleotides - metabolism; Oligonucleotides; Oligonucleotides - analysis; Phosphatases; Phosphoric Diester Hydrolases - metabolism; Phosphorus Isotopes; Polynucleotides - analysis; Templates, Genetic; Thymine Nucleotides - analysis
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.69.11.3448

The sequence of DNA base pairs adjacent to the phosphodiester bonds cleaved by the RI restriction endonuclease in unmodified DNA from coliphage λ has been determined. The 5′-terminal nucleotide labeled with32P and oligonucleotides up to the heptamer were analyzed from a pancreatic DNase digest. The following sequence of nucleotides adjacent to the RI break made in λ DNA was deduced from these data and from the 3′-dinucleotide sequence and nearest-neighbor analysis obtained from repair synthesis with the DNA polymerase of Rous sarcoma virus$ \matrix 5^{\prime}\cdots \cdot \text{A/T}_{\text{p}}\text{G}^{\downarrow}{}_{\text{p}}\text{A}_{\text{p}}\text{A}_{\text{p}}\text{T}_{\text{p}}\text{T}_{\text{p}}\text{C}_{\text{p}}\text{T/A}\cdots \cdot 3^{\prime} \\ 3^{\prime}\cdots \cdot \text{T/A}_{\text{p}}\text{C}_{\text{p}}\text{T}_{\text{p}}\text{T}_{\text{p}}\text{A}_{\text{p}}\text{A}_{\text{p}}{}_{\uparrow}\text{G}_{\text{p}}\text{A/T}\cdots \cdot 5^{\prime} \endmatrix $The RI endonuclease cleavage of the phosphodiester bonds (indicated by arrows) generates 5′-phosphoryls and short cohesive termini of four nucleotides,pApApTpT. The most striking feature of the sequence is its symmetry.