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By Paul D. (Editor); Boyer

ISBN-10: 0080865917

ISBN-13: 9780080865911

ISBN-10: 0121227146

ISBN-13: 9780121227142

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N . Arai and A. Kornberg, JBC 256, 5294 (1981). R. L. Low, K. Arai, and A. Kornberg, PNAS 78, 1436 (1981). D. W. Mosbaugh and R. R. Meyer, JBC 255, 10239 (1980). J . B. Flanegan and G. R. Greenbcrg, JBC 252, 3019 (1977). I . DNA POLYMERASES-A PERSPECTIVE 13 tifying the subunit structure of the polymerases and the genes that encode them. Remarkable techniques are provided by genetic engineering for amplifying the levels of these scarce enzymes: Novel methods are available for isolating enzymes, such as the use of monoclonal antibodies resulting from cell fusions; isotopic and staining methods, a hundredfold more sensitive, can be used for tracing the enzymes; and improved techniques are in hand for obtaining well-defined DNAs as template-primers for assay.

Since only a single triphosphate is labeled in all experiments, a correction is made to allow use of the ratio of rates of radioactive nucleotide incorporation (R::,). 4 = LP;:4 (16) This expression is incorporated into Eq. (15) to yield N4 = LN,/R::, (17) The theoretical relationship between N4 and R::, for calf thymus DNA is shown in Fig. 5. The value of R::, is given by R::, = Rx:4(Tx/T4) (18) where R,:, is the ratio of label incorporated in limited (x = I , 2 , or 3) as compared to complete ( x = 4) reactions and T,/T, is calculated from Eq.

Henningsen, PNAS 65, 168 (1970). 8. P. T. Englund, M. P. Deutscher, T. Jovin, R. B. Kelly, N. R. Cozzarelli, and A. Kornberg, CSHSQB 33, 1 (1968). 9. J. Gnffith, J. A. Huberman, and A. Kornberg, J M B 55, 209 (1971). 3. 4. 5. 6. 7. 17 2. DNA POLYMERASE I OF Escherichia coli group that can form a dissociable complex with HgZ+ (10) and can be carboxymethylated without loss of activity (5). As determined by atomic absorption spectroscopy, pol I contains one Zn2+atom per molecule (/I). As yet there is no direct evidence for the participation of this metal in the catalytic cycle of the enzyme.

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Nucleic Acids, Part A, 3rd Edition by Paul D. (Editor); Boyer

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