Purified thermostable DNA polymerase obtainable from thermococcus litoralis

US5322785A · kind A · utility

Assignee

• New England Biolabs, Inc. · US

Inventors

• Donald G. Comb · Attleboro, US
• Francine B. Perler · Brookline, US
• Rebecca Kucera · Hamilton, US
• William E. Jack · Wenham, US

Key dates

• Filing date: Apr 17, 1991
• Grant date: Jun 21, 1994
• Expiry date: Apr 17, 2011

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/1252
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

There is provided an extremely thermostable enzyme obtainable from Thermococcus litoralis. The thermostable enzyme has a molecular weight of about 90,000-95,000 daltons, a half-life of about 60 minutes at 100.degree. C. in the absence of stabilizer, and a half-life of about 95 minutes at 100.degree. C. in the presence of stabilizer, such as octoxynol (TRITON X-100) or bovine serum albumin. The thermostable enzyme possesses a 3'-5' proofreading exonuclease activity. The thermostable enzyme may be native or recombinant and may be used for second-strand cDNA synthesis in cDNA cloning, DNA sequencing, and DNA amplification.