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(12) United States Patent (10) Patent No.: US 6,555,349 B1 O'donnell (45) Date of Patent: Apr

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(12) United States Patent (10) Patent No.: US 6,555,349 B1 O'donnell (45) Date of Patent: Apr USOO6555349B1 (12) United States Patent (10) Patent No.: US 6,555,349 B1 O'Donnell (45) Date of Patent: Apr. 29, 2003 (54) METHODS FOR AMPLIFYING AND Xiao et al., “DNA Polymerase III Accessory Proteins. III. SEQUENCING NUCLEIC ACID MOLECULES HolC and holD Encoding X and up,” J. Biol. Chem. USING ATHREE COMPONENT 268: 11779–84 (1993). POLYMERASE Xiao et al., “DNA Polymerase III Accessory Proteins. IV. Characterization of X and up,” J. Biol. Chem. 268: 11773-78 (75) Inventor: Michael E. O'Donnell, (1993). Hastings-on-Hudson, NY (US) Studwell-Vaughan et al., “DNA Polymerase III Accessory (73) Assignees: Cornell Research Foundation, Inc., Proteins. V. 0 Encoded by holE,” J. Biol. Chem. Ithaca, NY (US); The Rockefeller 268:11785-91 (1993). University, New York, NY (US) Carter et al., “Molecular Cloning, Sequencing, and Overex pression of the Structural Gene Encoding the Ó Subunit of E. 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No. 5,668,004, which is a continuation-in-part of application No. 07/826, Carter et al., “Identification, Molecular Cloning and Char 926, filed on Jan. 22, 1993, now abandoned, application No. acterization of the Gene Encoding the X. Subunit of DNA 09/325,067, which is a continuation-in-part of application Polymerase III Holoenzyme of E. coli,' Mol. Gen. Genet. No. 09/282,917, filed on Mar. 31, 1999, now Pat. No. 6,221,642, which is a division of application No. 08/696, 241:399-408 (1993). 651, filed on Aug. 14, 1996, which is a continuation of Tomasiewicz et al., “Sequence Analysis of the Escherichia application No. 08/298.945, filed on Aug. 31, 1994, now Pat. No. 5,583,026. coli dnaE Gene,” J. Bacteriol. 169:5735–44 (1987). Ohmori et al., “Structural Analysis of the dnaA and dnaN (51) Int. Cl. ........................... C12Q 1/68; C12P 19/34; Genes of Escherichia coli,” Gene 28:159–170) (1984). 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