Open AccessRNA‐Dependent DNA Polymerase from a Cell Line Derived from the Bone Marrow of a Patient with Polycythemia vera
Author(s) -
WEIMANN Bernd Jürgen,
SCHMIDT Jörg,
KLUGE Norbert,
OSTERTAG Wolfram,
WOLFRUM Dirk Ingo
Publication year - 1975
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1975.tb02485.x
Subject(s) - microbiology and biotechnology , polymerase , centrifugation , ribonuclease , dna , gel electrophoresis , electrophoresis , primer (cosmetics) , polyacrylamide gel electrophoresis , chemistry , enzyme , polycythemia vera , dna polymerase , biochemistry , chromatography , rna , biology , gene , organic chemistry , immunology
A RNA‐dependent DNA polymerase was isolated from a human cell line derived from the bone marrow of a patient with polycythemia vera. The purification procedure included chromatography on phosphocellulose and oligo(dT)‐cellulose, and glycerol gradient centrifugation. The enzyme could be distinguished from polymerase A by salt elution from phosphocellulose, utilization of poly(rC) · oligo(dG) and its molecular size of about 70 000, as determined by centrifugation. Throughout the purification procedure ribonuclease H activity was co‐purified. Upon dodecyl‐sulfate‐polyacrylamide electrophoresis on microgradient gels two main bands with molecular weights of 68 000 and 66 000 and three minor bands were detected. The enzyme preferentially used poly(rA) · oligo(dT) as template‐primer compared with poly‐(dA) · oligo(dT). It incorporated dGMP into polymer on poly(rC) · oligo(dG).