Purification of Avian Myeloblastosis Virus DNA Polymerase by Affinity Chromatography on Polycytidylate-Agarose
- 1 October 1974
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 14 (4) , 853-9
- https://doi.org/10.1128/jvi.14.4.853-859.1974
Abstract
Polycytidylic acid [poly(rC)] covalently linked to cyanogen bromide-activated agarose is an effective affinity matrix for the RNA-dependent DNA polymerase from avian myeloblastosis virus. Poly(rC)-agarose is capable of binding large quantities of avian myeloblastosis DNA polymerase, which is then eluted by using a linear KCl gradient of increasing concentration. The DNA polymerase isolated from crude, detergent-disrupted virions by a single pass through columns of poly(rC)-agarose appears nearly homogeneous (approximately 90% pure) as determined by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis. Complete recovery of input enzymatic activity was obtained. Results suggest that polyribonucleotide columns may provide a high-yield, rapid method for the purification of oncornaviral DNA polymerase.Keywords
This publication has 20 references indexed in Scilit:
- Human DNA Polymerase III (R-DNA Polymerase): Distinction from DNA Polymerase I and Reverse TranscriptaseScience, 1974
- Evidence for template-specific sites in DNA polymerasesBiochemical and Biophysical Research Communications, 1974
- Utilization of mammalian 70S RNA by a purified reverse transcriptase from human myelocytic leukemic cellsBiochemical and Biophysical Research Communications, 1973
- Use of affinity matrices in determining steric requirements for substrate binding: Binding of anthranilate 5-phosphoribosylpyrophosphate phosphoribosyltransferase from Salmonella typhimurium to sepharose-anthranilate derivativesAnalytical Biochemistry, 1972
- Viral and Cellular DNA Polymerase: Comparison of Activities with Synthetic and Natural RNA TemplatesScience, 1972
- Affinity Chromatography of MacromoleculesPublished by Wiley ,1972
- Purification of the DNA polymerase of avian myeloblastosis virusBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1971
- Affinity ChromatographyAnnual Review of Biochemistry, 1971
- Covalent attachment of nucleic acids to agarose for affinity chromatographyBiochemistry, 1971
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970