Purification of RNA-directed DNA polymerase from mouse spleen infected with Rauscher leukemia virus.
RNA-directed DNA polymerase was purified from spleens of Balb/c and NMRI mice infected with Rauscher murine leukemia virus. The method includes cell fractionation and lysis of microsomal fraction, chromtography on Sephadex G-200 and phosphocellulose. Estimation of molecular weight from the sedimentation rate of the purified enzyme in a glycerol gradient was consistent with a structure containing one polypeptide with a molecular weight of 70,000. Purified RLV DNA polymerase from spleen could transcribe purified DNA polymerase from purified virions. This simple preparation method offers a procedure for large scale preparation of the RNA-directed DNA polymerase which can be used for synthesis of DNA complementary to mRNA.
D000818:Animals; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D011071:Poly T / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D011925:Rauscher Virus / Q000201:enzymology*; D013154:Spleen / Q000201:enzymology*
D011071:Poly T; D012194:RNA-Directed DNA Polymerase