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

Sensitive assay for mitochondrial DNA polymerase gamma.

R K Naviaux1, D Markusic, B A Barshop

  • 1The Mitochondrial and Metabolic Disease Center, University of California, San Diego, USA. naviaux@ucsd.edu

Clinical Chemistry
|October 3, 1999
PubMed
Summary

A new assay accurately measures mitochondrial DNA polymerase gamma activity, aiding diagnosis of mitochondrial DNA depletion syndromes. This enzyme is crucial for DNA replication, and its deficiency can cause severe genetic disorders.

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

Nicotine withdrawal-induced inattention is absent in alpha7 nAChR knockout mice.

Psychopharmacology·2017
Same author

Successful Domino Liver Transplantation from a Patient with Methylmalonic Acidemia.

JIMD reports·2015
Same author

Mitochondrial disorders affecting the nervous system.

Seminars in neurology·2014
Same author

Reversal of autism-like behaviors and metabolism in adult mice with single-dose antipurinergic therapy.

Translational psychiatry·2014
Same author

On being a Doctor... "and describe a disease not previously reported, Lesch--Nyhan disease".

Revista clinica espanola·2012
Same author

Severe hypertriglyceridemia and recurrent pancreatitis in a girl with type Ia glycogen storage disease and type III hyperlipoproteinemia.

American journal of medical genetics. Part A·2009

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Mitochondrial DNA polymerase gamma is essential for replicating mitochondrial DNA.
  • Deficiencies in this enzyme can lead to mitochondrial DNA depletion syndromes.
  • Existing diagnostic methods require enhancement for enzyme activity assessment.

Purpose of the Study:

  • To develop and validate a sensitive and robust clinical assay for mitochondrial DNA polymerase gamma activity.
  • To establish kinetic parameters and performance characteristics of the assay.
  • To evaluate the assay's utility in patients with suspected mitochondrial disorders.

Main Methods:

  • Mitochondria were isolated from skeletal muscle biopsies.
  • DNA polymerase gamma activity was measured using a substrate and captured on ion-exchange paper.

Related Experiment Videos

  • Products were quantified via autoradiography and scintillation counting.

    • Main Results:

    • The assay demonstrated high sensitivity with a detection limit of 30 units.
    • Kinetic parameters (Km for dTTP, Ki for azidothymidine triphosphate) were determined.
    • The assay showed good precision (6.4% within-day, 12% between-day) and a wide dynamic range.
    • Application to patient samples revealed significant biological variation in enzyme activity.

    Conclusions:

    • The developed assay is a valuable tool for evaluating patients with suspected mitochondrial DNA depletion.
    • It provides quantitative data on DNA polymerase gamma activity.
    • This assay complements existing diagnostic approaches for mitochondrial diseases.