Relaxed circular SV40 DNA as cleavage intermediate of two restriction endonucleases
- 1 January 1977
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 4 (6) , 1803-1814
- https://doi.org/10.1093/nar/4.6.1803
Abstract
We have determined the mode of cleavage of superhelical SV40 DNA (Form I) by restriction endonucleases EcoRI and HpaII at 37 degrees C. By analysis with agarose gel electrophoresis and direct examination with dark field electron microscopy, we found that a large amount of the single-nicked circular DNA (Form II) was produced before the linear SV40 DNA (Form III) appeared. Thus, both restriction enzymes cleave only one strand of the superhelical DNA first. The second cleavage on the complementary strand occurred after a lag period. The first order rate constant for the second cleavage by EcoRI endonuclease was determined and a kinetic reaction scheme for both enzymes is proposed.Keywords
This publication has 8 references indexed in Scilit:
- EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme.Journal of Biological Chemistry, 1976
- Nucleotide sequence analysis of DNA. XXIV. Synchronous digestion of SV40 DNA by exonuclease IIIBiochemistry, 1976
- Restriction and modification of a self-complementary octanucleotide containing the EcoRI substrateJournal of Molecular Biology, 1975
- Studies on the cleavage of bacteriophage lambda DNA with EcoRI restriction endonucleaseJournal of Molecular Biology, 1975
- Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresisBiochemistry, 1973
- DNA Nucleotide Sequence Restricted by the RI EndonucleaseProceedings of the National Academy of Sciences, 1972
- Cleavage of DNA by R 1 Restriction Endonuclease Generates Cohesive EndsProceedings of the National Academy of Sciences, 1972
- Specificity of the Break Produced by Restricting Endonuclease R 1 in Simian Virus 40 DNA, as Revealed by Partial Denaturation MappingProceedings of the National Academy of Sciences, 1972