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Separation of a New Deoxyribonucleic Acid Polymerase from Vaccinia-infected HeLa Cells

Separation of a New Deoxyribonucleic Acid Polymerase from Vaccinia-infected HeLa Cells K. I. Berns , C. Silverman and A. Weissbach 1 Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20014 ABSTRACT HeLa cell deoxyribonucleic acid (DNA) polymerase was purified about 100-fold by sequential column chromatography on phosphocellulose, hydroxylapatite, and Bio Rex 70. A new form of DNA polymerase found in vaccinia virus-infected cells was separated from HeLa DNA polymerase by chromatography on diethylamino-ethyl cellulose. The new form was also purified approximately 100-fold in the same manner as the HeLa DNA polymerase. In addition to chromatographic differences, the two enzymes differed with regard to primer response, relative activity at high p H, inactivation by heat and p -chloromercuribenzoate, and inhibition by vaccinia antiserum. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Virology American Society For Microbiology

Separation of a New Deoxyribonucleic Acid Polymerase from Vaccinia-infected HeLa Cells

Abstract

Separation of a New Deoxyribonucleic Acid Polymerase from Vaccinia-infected HeLa Cells K. I. Berns , C. Silverman and A. Weissbach 1 Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20014 ABSTRACT HeLa cell deoxyribonucleic acid (DNA) polymerase was purified about 100-fold by sequential column chromatography on phosphocellulose, hydroxylapatite, and Bio Rex 70. A new form of DNA polymerase found in vaccinia virus-infected cells was separated from HeLa DNA polymerase by chromatography on diethylamino-ethyl cellulose. The new form was also purified approximately 100-fold in the same manner as the HeLa DNA polymerase. In addition to chromatographic differences, the two enzymes differed with regard to primer response, relative activity at high p H, inactivation by heat and p -chloromercuribenzoate, and inhibition by vaccinia antiserum.
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