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Replication of phage phi 29 DNA with purified terminal protein and DNA polymerase: synthesis of full-length phi 29 DNA


, : Replication of phage phi 29 DNA with purified terminal protein and DNA polymerase: synthesis of full-length phi 29 DNA. Proceedings of the National Academy of Sciences of the United States of America 82(19): 6404-6408

A system that replicates bacteriophage phi 29 DNA with protein p3 covalently attached to the two 5' ends, using as the only proteins the phi 29 DNA polymerase and the terminal protein, is described. Restriction analysis of the 32P-labeled DNA synthesized in vitro showed that all phi 29 DNA fragments were labeled. Analysis by alkaline sucrose gradient centrifugation of the DNA labeled during a 10-min pulse showed that, after a 20-min chase, about half of the DNA molecules had reached apparently full-length phi 29 DNA (approximately equal to 18,000 nucleotides). Ammonium ions strongly stimulated phi 29 DNA-protein p3 replication, the effect being due to stimulation of the initiation reaction. ATP was not required for phi 29 DNA-protein p3 replication, either in the initiation or elongation steps. The results show that the phi 29 DNA polymerase functions, not only in the formation of the p3-dAMP covalent initiation complex but also in the elongation of the latter, as the only DNA polymerase to produce full-length phi 29 DNA.

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PMID: 3863101


Other references

van Heusden, G.P.; Noteborn, H.P.; ven Den Bosch, H., 1981: Selective utilization of palmitoyl lysophosphatidylcholine in this synthesis of disaturated phosphatidylcholine in rat lung: a combined in vitro and in vivo approach. 1. The acyl-CoA:lysophosphatidylcholine acyltransferase system in rat lung microsomes was found to utilize selectively 1-[1-14C]palmitoyl-sn-glycero-3-phosphocholine when compared with 1-[9,10-3H2]stearoyl-sn-glycero-3-phosphocholine. This result...

Stumper, R., 1923: Sur la composition chimique des nids de l'Apicotermes occultus Silv. Cd. Sci. Paris: 409-411

Sturkie, P.D.; Woods, J.J.; Meyer, D., 1972: Serotonin levels in blood, heart, and spleen of chickens, ducks, and pigeons. The content in spleens of fowls was very high compared to ducks (6.04-11.52), pigeons (2.80-5.40), and most mammals, and the concentration was higher in the female sex of all three species. Amounts in the heart of each species were not very differ...

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Harries, J.T.; Sladen, G.E., 1972: The effects of different bile salts on the absorption of fluid, electrolytes, and monosaccharides in the small intestine of the rat in vivo. Absorption was measured in fasting male Wistar rats by a closed-loop technique involving proximal jejunum and distal ileum. In jejunum, deoxycholate at a concentration of 1 mM impaired absorption of water and K but not of Na or glucose; at higher...

Beldeanu, E., 1974: Fruit production in Hippophae rhamnoides. Summarizes studies during 1969-71 near Cimpina in the same part of Rumania as those already noticed [see FA 35, 1407]. New suckers began to bear at 3 years old. Fruit production culminated at 7-8 years old, plants of a good clone yielding > 4 k...

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Bridge, S.; Christie, M.J.; Beart, P.M., 1986: No change in neostriatal D-2 dopamine receptors after NMDA lesions of rat prefrontal cortex. D-2 dopamine receptor function in rat nucleus accumbens and anterior corpus striatum was examined 6-8 days following N-methyl-D-aspartate lesions confined to medial prefrontal cortex. Lesions failed to alter the affinity or density of D-2 receptor...

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