Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

EcoRV restriction endonuclease binds all DNA sequences with equal affinity.

J D Taylor1, I G Badcoe, A R Clarke

  • 1Department of Biochemistry, University of Bristol, U.K.

Biochemistry
|September 10, 1991
PubMed
Summary

The EcoRV restriction enzyme binds non-specifically to all DNA sequences without Mg2+. In the presence of Mg2+, the enzyme translocates to its specific recognition site for cleavage.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Clinicopathological findings of melioidosis in captive red-legged pademelons (Thylogale stigmatica) in northern Queensland, Australia.

Australian veterinary journal·2024
Same author

Toxoplasmosis epidemic in a population of urbanised allied rock-wallabies (Petrogale assimilis) on Magnetic Island (Yunbenun), North Queensland.

Australian veterinary journal·2024
Same author

Correction: Functional redundancy between Apc and Apc2 regulates tissue homeostasis and prevents tumorigenesis in murine mammary epithelium.

Oncogene·2024
Same author

A novel alphaherpesvirus and concurrent respiratory cryptococcosis in a captive koala (Phascolarctos cinereus).

Australian veterinary journal·2022
Same author

A comprehensive phylogeny helps clarify the evolutionary history of host breadth and lure response in the Australian Dacini fruit flies (Diptera: Tephritidae).

Molecular phylogenetics and evolution·2022
Same author

The contraceptive efficacy of a self-assembling intra-uterine device in domestic mares.

Australian veterinary journal·2021

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Enzymology

Background:

  • EcoRV restriction endonuclease is known for highly specific DNA cleavage in the presence of MgCl2.
  • The mechanism of EcoRV binding to DNA, particularly in the absence of Mg2+, requires further investigation.

Purpose of the Study:

  • To investigate the binding of EcoRV restriction enzyme to DNA in the absence of Mg2+.
  • To characterize the nature of DNA-protein complexes formed.
  • To develop a method for analyzing multi-site protein binding on DNA.

Main Methods:

  • Polyacrylamide gel electrophoresis to detect DNA-protein complexes.
  • Analysis of DNA fragments of varying lengths (55 bp and 381 bp).
  • Development of a novel method for analyzing protein binding to multiple DNA sites.

Related Experiment Videos

Main Results:

  • EcoRV binds to DNA non-specifically in the absence of Mg2+, forming multiple complexes dependent on DNA length.
  • No difference in binding was observed between DNA with or without the EcoRV recognition site.
  • A new method revealed that EcoRV binds all sequences with the same equilibrium constant, but exhibits cooperative binding at adjacent sites.
  • Addition of MgCl2 induced translocation of EcoRV from non-specific to specific sites.

Conclusions:

  • EcoRV exhibits non-specific DNA binding in the absence of Mg2+.
  • Cooperative binding of EcoRV to adjacent sites occurs.
  • Mg2+ is crucial for directing EcoRV to its specific recognition site, facilitating cleavage through translocation.