Physical Characterization and Simultaneous Purification of Bacteriophage T4 Induced Polynucleotide Kinase, Polynucleotide Ligase, and Deoxyribonucleic Acid Polymerase

A. Panet, J. H. van de Sande*, P. C. Loewen, H. G. Khorana, A. J. Raae, J. R. Lillehaug, K. Kleppe

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

224 Scopus citations

Abstract

The initial steps in the existing procedures for the purification of the bacteriophage T4 induced polynucleotide kinase, ligase, and the DNA polymerase have been modified so as to enable their simultaneous preparation from one batch of the infected Escherichia coli cells. Procedures were devised to remove an exonuclease which often contaminated the preparations of the kinase and ligase. The kinase was shown to be homogeneous by gel electrophoresis. Its molecular weight under native conditions using gel filtration was found to be 140,003 ± 10% daltons. Under denaturing conditions employing sodium dodecyl sulfate electrophoresis the mol wt was estimated to be 33,000 ±5% daltons. The ligase also gave a single band upon gel electrophoresis and the mol wt under denaturing conditions was 63,000 ± 5 % daltons. Gel filtration of the latter enzyme in the absence of denaturing agents gave a mol wt of 68,000 ± 10% daltons.

Original languageEnglish
Pages (from-to)5045-5050
Number of pages6
JournalBiochemistry
Volume12
Issue number25
DOIs
StatePublished - 1 Dec 1973
Externally publishedYes

Fingerprint

Dive into the research topics of 'Physical Characterization and Simultaneous Purification of Bacteriophage T4 Induced Polynucleotide Kinase, Polynucleotide Ligase, and Deoxyribonucleic Acid Polymerase'. Together they form a unique fingerprint.

Cite this