Expression of an enzymatically active murine retroviral reverse transcriptase in human cells

• Published in: Biochemical and biophysical research communications Vol. 158; no. 2; pp. 595 - 602
• Main Authors: Jean-Jean, Olivier, Moyret, Caroline, Bernard, Dominique, de Recondo, Anne-Marie, Rossignol, Jean-Michel
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 31.01.1989; Elsevier
• Subjects: Animals; Biological and medical sciences; Cells, Cultured; Cloning, Molecular; Endoribonucleases - genetics; Endoribonucleases - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation; Genetic Engineering; Humans; Immunologic Techniques; Mice; Molecular and cellular biology; Molecular genetics; Plasmids; Regulatory Sequences, Nucleic Acid; Ribonuclease H; RNA-Directed DNA Polymerase - genetics; RNA-Directed DNA Polymerase - immunology; RNA-Directed DNA Polymerase - metabolism; Time Factors; Human; Cell culture; Oncovirinae; Radiolabelling; Enzyme; Retroviridae; Gene expression; Virus; Proteins; Cell line; Transfection; Enzymatic activity; Gene; Type C oncovirus; DNA polymerase RNA-dependent; Immunoblotting assay; Murine leukemia virus; Inhibition; Vector; Nucleic acid
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/S0006-291X(89)80091-9

The region of the pol gene of the Moloney murine leukemia virus (M-MuLV) encoding the reverse transcriptase and RNase H activities was inserted in an eukaryotic expression vector and transiently expressed in human cultured cells. This results in the expression of high levels of reverse transcriptase activity. This enzyme, partially purified, also carries a RNase H activity, has the biochemical requirements of the viral enzyme and is recognized and inhibited by antibodies directed against a M-MuLV reverse transcriptase expressed in Escherichia coli.