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

Phosphatases and kinases.

S Tabor1

  • 1Harvard Medical School, Boston, Massachusetts, USA.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary
This summary is machine-generated.

This study details bacterial alkaline phosphatase (BAP) and calf intestine phosphatase (CIP) for nucleic acid dephosphorylation. It also covers T4 polynucleotide kinase for radioactive labeling and phosphate exchange in molecular biology.

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

Asymmetric split H-shape nanoantennas for molecular sensing.

Biomedical optics express·2017
Same author

The impact of feeding diets of high or low energy concentration on carcass measurements and the weight of primal and subprimal lean cuts.

Asian-Australasian journal of animal sciences·2014
Same author

Enzymatic labeling of DNA.

Current protocols in human genetics·2008
Same author

Expression using the T7 RNA polymerase/promoter system.

Current protocols in molecular biology·2008
Same author

Exonucleases.

Current protocols in molecular biology·2008
Same author

Endonucleases.

Current protocols in molecular biology·2008
Same journal

Nondenaturing Polyacrylamide Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Purification and Concentration of DNA from Aqueous Solutions: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Expression of Proteins Using Semliki Forest Virus Vectors: Protein Expression.

Current protocols in molecular biology·2021
Same journal

Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions: DNA-Protein Interactions.

Current protocols in molecular biology·2021
Same journal

Separation of Double- and Single-Stranded Nucleic Acids Using Hydroxylapatite Chromatography: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Pulsed-Field Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
See all related articles

Area of Science:

  • Molecular Biology
  • Enzymology

Background:

  • Bacterial alkaline phosphatase (BAP) and calf intestine phosphatase (CIP) are key enzymes in nucleic acid research.
  • These enzymes remove 5'-phosphate groups from DNA, RNA, and nucleotide triphosphates.

Purpose of the Study:

  • To describe the reaction conditions and applications of BAP, CIP, and T4 polynucleotide kinase.
  • To explain the use of these enzymes in radioactive labeling and phosphate modification of nucleic acids.

Main Methods:

  • Enzymatic hydrolysis of 5'-phosphate residues using BAP and CIP.
  • Radioactive labeling of 5'-hydroxyl termini with [gamma-32P]ATP using T4 polynucleotide kinase.
  • Characterization of T4 polynucleotide kinase activities: forward phosphorylation, exchange reaction, and 3' phosphatase activity.

Related Experiment Videos

Main Results:

  • BAP and CIP effectively dephosphorylate various nucleic acid and nucleotide substrates.
  • Dephosphorylated nucleic acids are suitable for subsequent radioactive labeling via T4 polynucleotide kinase.
  • T4 polynucleotide kinase exhibits efficient forward phosphorylation, phosphate exchange, and 3' phosphatase activities.

Conclusions:

  • BAP and CIP are essential tools for preparing nucleic acids for labeling and further manipulation.
  • T4 polynucleotide kinase is versatile, enabling radioactive labeling, phosphate exchange, and dephosphorylation.
  • These enzymes collectively facilitate advanced molecular biology techniques in nucleic acid research.