[RNA-dependent DNA polymerase from rat liver].
RNA-dependent DNA-polymerase was isolated from rat liver, and its characteristics were studied. Wistar rat livers were homogenized in the disruptive buffer, centrifuged at 100,000 g and the supernatant was freed of the nucleic acids by DEAE-cellulose (DE-23) chromatography. The further chromatography of the eluate on DEAE-cellulose (DE-52) and phosphocellulose P-11 resulted in the obtaining of 300-400-fold purified RNA-dependent DNA-polymerase. Even more purified enzyme 1500-fold was isolated from the 165,000 g pellet of postmitochondrial rat liver fraction. The main properties of the purified enzyme are characteristic for the retroviral reverse transcriptase. The enzyme catalyzes DNA synthesis when poly(A)+mRNA is used as a template-primer. Its sedimentation constant amounts to 4.6 S. Mg2+ is preferable to Mn2+ as an activator of the enzyme. The optimal pH is 7.8. Among the products of the enzymic reaction hybrid RNA X DNA molecules were identified.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D006498:Hepatectomy; D008099:Liver / Q000201:enzymology* / Q000382:microbiology; D008862:Microsomes, Liver / Q000201:enzymology; D012194:RNA-Directed DNA Polymerase / Q000032:analysis* / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D012190:Retroviridae / Q000201:enzymology; D014771:Virion / Q000201:enzymology
D012194:RNA-Directed DNA Polymerase
RNK-zavisimaia DNK-polimeraza iz pecheni krys.