Purification of avian myeloblastosis virus DNA polymerase by affinity chromatography on polycytidylate-agarose.
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.
D001354:Avian Leukosis Virus / Q000201:enzymology*; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003597:Cytosine Nucleotides; D004591:Electrophoresis, Polyacrylamide Gel; D006160:Guanosine Triphosphate / Q000378:metabolism; D008970:Molecular Weight; D011119:Polynucleotides; D011134:Polysaccharides; D011189:Potassium Chloride; D012194:RNA-Directed DNA Polymerase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism; D014316:Tritium
D003597:Cytosine Nucleotides; D011119:Polynucleotides; D011134:Polysaccharides; D013942:Thymine Nucleotides; D014316:Tritium; D011189:Potassium Chloride; D006160:Guanosine Triphosphate; D012194:RNA-Directed DNA Polymerase