Separation of cellular and viral DNA polymerases by affinity chromatography on polynucleotide-Sepharose.
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.
D000818:Animals; D000937:Antigen-Antibody Reactions; D002846:Chromatography, Affinity / Q000379:methods; D004273:DNA, Neoplasm / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D007942:Leukemia, Experimental / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011068:Poly G; D011072:Poly U; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism; D011925:Rauscher Virus / Q000378:metabolism; D013154:Spleen / Q000378:metabolism
D004273:DNA, Neoplasm; D004279:DNA, Viral; D011068:Poly G; D011072:Poly U; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase