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Separation of cellular and viral DNA polymerases by affinity chromatography on polynucleotide--Sepharose

Journal Article · · Cancer Biochem. Biophys.; (United States)
OSTI ID:7091147
Polyguanylate- and poly(2'-O-methyl)uridylate-Sepharose have been prepared for affinity chromatography of DNA polymerases of viral origin (reverse transcriptase). Both cellular DNA polymerases and reverse transcriptase bind to polyguanylate-Sepharose. The cellular polymerases can be eluted from the column between 0.32 and 0.42 M NaCl while reverse transcriptase eluted between 0.56 and 0.78 M NaCl. However, only reverse transcriptase adheres to poly(2'-O-methyl)uridylate-Sepharose and can be eluted at approximately 0.35 M NaCl. The columns were used to partially purify RNA-dependent DNA polymerase from spleens of mice infected with Rauscher leukemia virus. The enzyme preparation is about 1300-fold purified and is inhibited by antiserum prepared against purified reverse transcriptase from Rauscher leukemia virus to the same extent as the virion enzyme.
Research Organization:
Oak Ridge National Lab., TN
OSTI ID:
7091147
Journal Information:
Cancer Biochem. Biophys.; (United States), Journal Name: Cancer Biochem. Biophys.; (United States) Vol. 1; ISSN CABCD
Country of Publication:
United States
Language:
English

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