Method of synthesizing polydeoxyribonucleotides

Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives

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435 6, 435 911, 435 912, 435 915, 536 231, 536 243, 536 241, 536 242, C07H 2100, C07H 2102, C07H 2104, C12P 1934

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059170310

ABSTRACT:
The invention relates to a method of synthesizing polydeoxyribonucleotides which comprises causing a thermostable deoxyribunucleotide polymerase to act on deoxyribonucleotides without using any template and any primers, to the thus-obtained polydeoxyribonucleotides, to a method of screening cDNA libraries using the same as probes, to a method comprising joining the DNA resulting from the above method of synthesis to the 3'--OH terminus of a double-stranded DNA containing a gene derived from cells of a living organism, followed by transfection of such cells with the product of joining and culture of the cells, to thereby cause insertion of the product into chromosomal DNA and production of the protein encoded by the gene, to a method of synthesizing chromosomal DNA and to a method of synthesizing chromosomes. According to the invention, it is possible to create DNA not occurring in the nature and create novel proteins using the thus created genes, hence means for providing novel physiologically or pharmacologically active substances can be provided.

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Henner et al, "De novo synthesis of a polymer of deoxyadenylate and deoxythymidylate by calf thymus DNA polymerase alpha", Proc. Natl. Acad. Sci. 72(10):3944-3946, Oct. 1975.
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Hanaki et al. "Primer/template independent synthesis of poly d(A-T) by Taq polymerase", Biochem. Biophys. Res. Comm. 238:113-118, 1997.
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Enzymatic Synthesis of Deoxyribonucleic Acid XII. A Polymer of Deoxyguanylate and Deoxyoytidylate, Charles M. Radding, John Josse and Arthur Kornberg, The Journal of Biological Chemistry, vol. 237, No. 9, Sep. 1962.
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Effect of Incubation Conditions on the Nucleotide Sequence of DNA Products of Unprimed DNA Polymerase Reactions, John F. Burd and Robert D. Wells, J. Mol. Biol. (1970) 53, 435.

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