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DNA polymerases in human lymphoblastoid cells infected with simian sarcoma virus

1974; Elsevier BV; Volume: 349; Issue: 2 Linguagem: Inglês

10.1016/0005-2787(74)90076-8

1879-3002

B.J. Lewis, John W. Abrell, R. Guy Smith, Robert C. Gallo,

Virus-based gene therapy research

Abstract This report describes a technique for isolating and separating DNA polymerases in mammalian cells infected with an RNA tumor virus. In whole cell extracts of cultured human lymphoblastoid cells infected with simian sarcoma virus, four DNA polymerases were resolved. Three of the enzymes were cellular, and one was viral in origin. The viral reverse transcriptase and cellular DNA polymerase II were eluted from a DEAE-cellulose column at 50 mM KCl, while cellular DNA polymerases I and III (R-DNA polymerase) were removed by 0.3 M KCl. These DNA polymerase mixtures were then resolved on phosphocellulose and DNA-cellulose columns. The identity of the separate polymerases was further confirmed by primer template utilization, molecular size determination, and response to antibody to the viral enzyme. This procedure was designed for further application to the study of DNA polymerases in human leukemic cells and other human tumors.