Kung, Patrick C. and Gottlieb, Paul D. and Baltimore, David (1976) Terminal deoxynucleotidyltransferase. Serological studies and radioimmunoassay. Journal of Biological Chemistry, 251 (8). pp. 2399-2404. ISSN 0021-9258. http://resolver.caltech.edu/CaltechAUTHORS:KUNjbc76
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Mouse antisera against calf terminal deoxynucleotidyltransferase (terminal transferase) have been prepared. The sera have been used to characterize terminal transferase both by studying inhibition of enzyme activity and by developing a competition radioimmunoassay using highly purified 125I-labeled terminal transferase. By either assay, anti-terminal transferase serum did not cross-react significantly with calf DNA polymerases alpha and beta, Escherichia coli DNA polymerase I, or the reverse transcriptase of Moloney mouse leukemia virus. The calf terminal transferase did, however, share cross-reactive but not identical determinants with human and murine terminal transferase. The radioimmunoassay could detect as little as 2 ng of terminal transferase/mg of soluble protein in a tissue extract. Thymocytes were found to contain 280 ng of terminal transferase/mg of cell protein or about 1 X 10^(5) molecules/cell; bone marrow had about 1% of the level of enzyme found in thymus. Extracts of spleen, peripheral white blood cells, lymph nodes, liver, muscle, and kidney all lacked detectable antigenicity of terminal transferase. These data indicate that terminal transferase is a tissue-specific enzyme and is not related to other DNA polymerases.
|Additional Information:||Copyright © 1976 by the American Society for Biochemistry and Molecular Biology. (Received for publication, October 28, 1975) We are very grateful to Dr. L.M.S. Chang for the gifts of calf DNA polymerases α and β, and for her advice on the purification of terminal deoxynucleotidyl-transferase from the calf thymus. Thanks are also extended to Drs. Ronald P. McCaffrey, Robert L. Ratliff, Allen Silverstone, and Amos Panet and Mr. Thomas A. Harrison for their stimulating discussion and supplies of some material. This work was supported by Grants CA-14051 and CA-15808 from the National Institutes of Health and by a contract from the Virus Cancer Program of the National Cancer Institute. [P.C.K. was a] Postdoctoral Fellow of the Jane Coffin Childs Fund for Medical Research. [D.B. was an] American Cancer Society Professor of Microbiology.|
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