Heat-Labile Alkaline Phosphatase from Antarctic Bacteria: Rapid 5' End-Labeling of Nucleic Acids
Abstract
A heat-labile alkaline phosphatase has been purified to near homogeneity from HK47, a bacterial strain isolated from Antarctic seawater. The active form of the enzyme has a molecular weight of 68,000 and is uniquely monomeric. The optimal temperature for the enzymatic activity is 25°C. Complete and irreversible thermal inactivation of the enzyme occurs in 10 min at 55°C. By using this heat-labile enzyme for dephosphorylation followed by a 10-min heat treatment, rapid end-labeling of nucleic acids by T4 polynucleotide kinase has been achieved.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- November 1984
- DOI:
- 10.1073/pnas.81.21.6691
- Bibcode:
- 1984PNAS...81.6691K