What question did this study set out to answer?

To characterize the mechanism of DNA cleavage by R1 restriction endonuclease and evaluate whether the resulting DNA ends can be rejoined to produce viable or recombinant molecules.

November 1, 1972Proceedings of the National Academy of SciencesOpen Access

Cleavage of DNA by R 1 Restriction Endonuclease Generates Cohesive Ends

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Authors

JMJanet E. MertzUniversity of Wisconsin–Madison RDRonald W. DavisWestern Carolina University

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Implication

In vitro biochemical analysis demonstrates R1 endonuclease cleaves DNA into complementary cohesive ends joinable by ligase, suggesting a universal mechanism for recombinant DNA.

Key Points

To characterize the mechanism of DNA cleavage by R1 restriction endonuclease and evaluate whether the resulting DNA ends can be rejoined to produce viable or recombinant molecules.
Digested duplex SV40 DNA with purified R1 restriction endonuclease.
Analyzed the chemical termini and sequence symmetry of the cleaved single-stranded ends.
Incubated cleaved linear SV40 DNA fragments with Escherichia coli DNA ligase to test circularization and infectivity.
R1 endonuclease makes staggered single-strand breaks producing identical, self-complementary single-stranded ends with 5'-phosphoryl and 3'-hydroxyl termini.
E. coli DNA ligase successfully rejoins cleaved linear SV40 DNA into fully infectious, covalently closed circular duplex molecules.
All R1-generated termini are identical and complementary, allowing any two DNA fragments cleaved by R1 to be joined together into hybrid molecules.

Cite This Study

Mertz et al. (1972) studied this question.

synapsesocial.com/papers/6a1d233f102421609404e906 https://doi.org/10.1073/pnas.69.11.3370

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Implication

In vitro biochemical analysis demonstrates R1 endonuclease cleaves DNA into complementary cohesive ends joinable by ligase, suggesting a universal mechanism for recombinant DNA.

Cleavage of DNA by R 1 Restriction Endonuclease Generates Cohesive Ends

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Implication

Key Points

Cite This Study

Cleavage of DNA by R 1 Restriction Endonuclease Generates Cohesive Ends

Discussion

Member takes

Implication

Key Points

Cite This Study

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