Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1982-11-24
1985-06-04
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 91, 4351723, 435317, 436 63, 436 94, 536 27, 935 17, 935 19, 935 77, C12Q 168, C12P 1934, C12N 1500, C12N 100, G01N 3114, C07H 2102, C07H 2104
Patent
active
045215090
ABSTRACT:
2'-deoxyadenosine 5'-0-(1-thiotriphosphate) (dATP(.alpha.-S)) was introduced into the 3'-ends of DNA restriction fragments with E. coli DNA polymerase I to give phosphorothioate internucleotide linkages. Such "capped " 3'-ends were found to be resistant to exonuclease III digestion. Moreover, the resistance to digestion is great enough that, under the conditions employed, just one strand of a double helix was digested by exonuclease III when "cap" was placed at only one end; when digestion was carried to completion, the production of intact single strands resulted. When digestion with exonuclease III was limited, and followed by S1 nuclease treatment, double stranded DNA fragments asymmetrically shortened from just one side were produced. In this way thousands of nucleotides can be selectively removed from one end of a restriction fragment. In vitro introduction of phosphorothioate linkages into one end of linearized replicative plasmid, followed by exonuclease III and S1 nuclease treatments, gave rise to truncated forms which, upon circularization by blunt end ligation, transformed E. coli and replicated in vivo.
REFERENCES:
Green et al.: Proc. Natl. Acad. Sci. USA, 77, 2455 (1980).
Weaver et al.: Nucleic Acids. Res., 7, 1175 (1979).
Kunkel et al., Proc. Natl. Acad. Sci., 78, 6734 (1981).
Bryant et al., Chem. Abstr., 94, 26753p (1981) of Phosphorus Chem. Directed Biol. Lect. Int. Symp., 1979, Stec (ed.), Pergamon, New York, pp. 129-131.
Shalloway et al., Cell 20, 411 (1980).
Orr, George A., et al., "Adenosine 5'-O-([.gamma.-.sup.18 0].gamma.-thio) Triphosphate Chiral at the .gamma.-phosphorus: Stereochemical Consequence of Reactions Catalyzed by Pyruvate Kinase, Glycerol Kinase, and Hexokinase", Proc. Natl. Acad. Sci., USA, vol. 75, No. 5, pp. 2230-2233, May 1978.
Kwan-Fu Rex Sheu, Frey, Perry A., "UDP-glucose Pyrophosphorylase", Stereochemical Course of the Reaction of Glucose 1-phosphate with Uridine-5'[1-thiotriphosphate] (1978) J. Biol. Chem., 253, 3378.
Atkinson, Maurice R., et al., "Enzymatic Synthesis of Deoxyribonucleic Acid XXXIV Termination of Chain Growth by a 2',3'-Dideoxyribonucleotide" (1969) Biochemistry 8, 4897-4904.
Sanger, F., Nicklen, S., Coulson, A. R.; "DNA Sequencing with Chain-Terminating Inhibitors", Proc. Natl. Acad. Sci., USA, vol. 74, pp. 5463-5467, Dec. 1977.
Putney, Scott D.; Benkovic, Stephen J., Schimmel, Paul R.; "A DNA Fragment with an .alpha.-phosphorothioate Nucleotide at One End is Asymmetrically Blocked from Digestion by Exonuclease III and can be Replicated In Vivo", Proc. Natl. Acad. Sci. USA, vol. 78, pp. 7350-7354, Dec. 1981.
Bryant et al.: Phosphorous Chem. Directed Biol. Lect. Int. Symp., Stec (ed.), Pergamon Press, New York (1979), pp. 129-131.
Burgers, Peter M. J., Eckstein, Fritz, "A Study of the Mechanism of DNA Polymerase I From Escherichia coli with Diastereomeric Phosphorothioate Analogs of Deoxyadenosine Triphosphate" (1979) J. Biol. Chem., 254, 6889.
Bryant, Floyd R., Benkovic, Stephen; "Stereochemical Course of the Reaction Catalyzed by 5'-Nucleotide Phosphodiesterase from Snake Venom" (1979) Am. Chem. Soc., p. 2825.
Eckstein, F., Armstrond, V. W., Steinbach, H.; "Stereochemistry of Polymerization by DNA-dependent RNA-polymerase from Escherichia coli: An Investigation with a Diastereomeric ATP-analogue", Proc. Natl. Acad. Sci., USA, vol. 73, No. 9, pp. 2987-2990, Sep. 1976.
Benkovic Stephen J.
Putney Scott D.
Schimmel Paul R.
Martinell James
Research Corporation
Wiseman Thomas G.
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