Clifford L. Harvey

Small-scale preparation of 5′-nucleotides and analogs by carrot phosphotransferase

Abstract Carrot phosphotransferase has been used to prepare a variety of 5′-nucleotides and analogs for E. coli DNA polymerase binding site studies. The ability of the enzyme to ignore a wide variety of structural variation and stereochemical detail, while preserving its specificity in substrate phosphorylation, makes it a useful tool in such preparations. The activity has been further characterized by kinetic studies.

Formation of poly (dA)-poly(dT) by Escherichia coli DNA polymerase in the presence of anthracycline antibiotics.

Abstract The antibiotics nogalamycin and ruticulomycin inhibited the primed or unprimed synthesis by Escherichia coli DNA polymerase of poly[d(A-T)] · poly[d(T-A)]. These antibiotics had no effect on the synthesis of the corresponding homopolymer poly(dA) · poly(dT). In the presence of nogalamycin, Escherichia coli DNA polymerase synthesized the homopolymer de novo .

High pressure liquid chromatography of oligodeoxyribonucleotides.

Abstract A high pressure liquid chromatography system capable of resolving oligodeoxy-ribonucleotides of intermediate size is described. Columns filled with a pellicular weak anion exchanger consisting of a polymeric aliphatic amine permit the rapid resolution of small amounts of oligomers. The method was applied to monitor a chemical condensation mixture leading to the tridecamer d-EtSp(T-T-A-G-C-A-G-C-C-G-C-A-G) ★ , and to assay an enzymatic ligation leading to the nonadecamer d(A-A-G-A-C-A-G-C-A-T-A-T-G-G-A-T-T-A-T). Both oligomers are constituents of a DNA duplex coding for S-Peptide2–14 of Ribonuclease A.