PURIFICATION OF RNA-DIRECTED DNA-POLYMERASE FROM MOUSE SPLEEN INFECTED WITH RAUSCHER LEUKEMIA-VIRUS

  • 1 January 1978
    • journal article
    • research article
    • Vol. 37  (11-1) , 1655-1663
Abstract
RNA-directed DNA polymerase [EC 2.7.7.7] was purified from spleens of Balb/c and NMRI mice infected with Rauscher murine leukemia virus [RLV]. The method includes cell fractionation and lysis of microsomal fraction, chromatography on Sephadex G-200 and phosphocellulose. Estimation of MW from the sedimentation rate of the purified enzyme in a glycerol gradient was consistent with a structure containing 1 polypeptide with a MW of 70,000. Purified RLV DNA polymerase from spleen could transcribe ribopolymers, deoxyribopolymers and heteropolymers as efficiently as described for 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.