By F. J. Bollum (auth.), Umberto Bertazzoni, F. J. Bollum (eds.)
This e-book comprises the court cases of a convention dedicated to the learn of the constitution and serve as of Terminal deoxynucleotidyl Transferase (TdT) and its usage as biochemical marker in immuno biology and leukemia, held in Elba, Italy on could 28-31, 1981. The en zyme has been identified to nucleic acid biochemists for greater than two decades and has proved to be a good device for making deoxypolymers, label ing DNA fragments, and including homopolymer tails to limit endonu clease fragments from DNA. because the discovery of its strange tissue distribution, quite often limited to the thymus and bone marrow, and of its irregular take place rence in human leukemic lymphoblasts, TdT has turn into essentially the most conventional markers within the examine of lymphocyte differentiation and within the type of hematopoietic neoplasia. the topic appeared appropriate for a gathering the place molecular and mobile biologists, immunologists and hematologists might convene for the 1st time to debate either uncomplicated study and the medical features of the matter. one of the targets accomplished through this workshop was once the sharing of in formation concerning the enzymology of TdT, biochemical and immunological technique and the correlation of TdT with different markers within the diag nosis of leukemia. The extraordinary accordance of effects, provided the following via 5 autonomous hematological associations, from research of TdT in different hundreds of thousands of leukemic sufferers, marks the significance of this enzyme as a diagnostic and prognostic instrument in those diseases.
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Sample text
SDS gel electrophoresis of human terminal transferase from blasts of some patients with chronic myelogenous leukemia in blast crisis and from all samples of normal human thymus. 46 M. R. , ET AL. 6700(}- 45000--- 3500(}- 1200(}- Figure 6. ferase. SDS gel electrophoresis of calf thymus terminal trans- Because the SDS gel patterns of the ~ = 62,000 form of the human terminal transferase were so different from the traditional two subunit structure characteristic of calf thymus terminal transferase 2 , properties of the native form of the enzyme from human blasts were investigated.
A gel identical to that shown in Fig. 2 was used for analysis of TdT peptides by the immunoblot procedure. Immunoreactive peptides were visualized by immunoperoxidase staining. The purification procedure described starts with frozen calf thymus glands. Low molecular weight immunoreactive peptides can be detected in the crude extract. It is not known at the present whether these low molecular weight forms of the enzyme have any biological significance. It is possible that the lower molecular weight forms arise during storage of the glands, although these forms are also found in extracts made from fresh tissue.
R. Deibel, Jr. S. Chem. 254: 8634 (1979). 6. A. Silverstone, L. N. Witte, and D. Bio1. Chem. 255: 791 (1980) 7. D. R. Commun. 72: 840 (1976). 8. H. Nakamura, K. Tanabe, S. Yoshida, and T. Chem. 256: in press (1981). 9. H. Nakamura, T. Morita, and S. Res. 38: 499 (1980). 10. K. Tanabe, M. Yamaguchi, A. Matsukage, and T. Bio1. Chem. 256: 3098 (1981). STRUCTURE OF CALF THYMUS TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE Lucy M. S. Chang and F. J. Bollum Uniformed Services University of the Health Sciences 4301 Jones Bridge Road, Bethesda, MD 20814 INTRODUCTION Terminal deoxynucleotidyl transferase (TdT), originally purified from calf thymus glands to homogeneity, was shown to have a native molecular weight of 32,000 by equilibrium sedimentation and two peptides of 26,500-dalton and 8,000-dalton when analyzed by gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) (1).