Purification and characterization of soluble and membrane-associated DNA polymerases from a virus PBCV-1 infected green alga Chlorella NC64A

Jan Rohozinski, James L. Van Etten

Research output: Contribution to journalArticle

Abstract

DNA polymerases were purified several hundred-fold from the 10 000 × g soluble (polymerase I) and particulate (polymerase III) fractions prepared from virus PBCV-1 infected Chlorella NC64A extracts. Both DNA polymerases exhibited optimal activity with activated calf thymus DNA at pH 8.5. DNA polymerase I required 3.0 mol m-3 MgSO4 and 150 to 250 mol m-3 KCl for optimum activity whereas, DNA polymerase III required 2.0 mol m-3 MgSO4 and 150 mol m-3 KCl. Both enzymes were inhibited by pyrophosphate, actinomycin D, ethidium bromide, dideoxythymidine triphosphate, and N-ethylmaleimide but were relatively insensitive to aphidicolin. DNA polymerase I differed from DNA polymerase III in its response to cations (particularly NH4Cl), elution from a DEAE cellulose column, and molecular weight.

Original languageEnglish (US)
Pages (from-to)1293-1298
Number of pages6
JournalJournal of experimental botany
Volume40
Issue number12
DOIs
StatePublished - Dec 1989

Keywords

  • Algal virus
  • Chlorella
  • DNA polymerase

ASJC Scopus subject areas

  • Physiology
  • Plant Science

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