Purified thermostable nucleic acid polymerase enzyme from thermosipho africanus

• US 5,968,799 A
• Filed: 02/09/1993
• Issued: 10/19/1999
• Est. Priority Date: 09/28/1990
• Status: Expired due to Term

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Greenfield, I. Lawrence, Reichert, Fred L., Gelfand, David H.
• Primary Examiner(s): WAX, ROBERT A
• Application Number:
  US07/977,428
• Time in Patent Office: 2,443 Days
• Field of Search: 435/194, 935/10, 935/14
• US Class Current: 435/194
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Abstract

A purified thermostable enzyme is derived from the eubacterium Thermosipho africanus. The enzyme has DNA polymerase, activity reverse transcriptase activity, and optionally 5'"'"'→3'"'"' and/or 3'"'"'→5'"'"' exonuclease activity. The enzyme can be native or recombinant, and may be used with primers and nucleoside triphosphates in a temperature-cycling chain reaction where at least one nucleic acid sequence is amplified in quantity from an existing sequence.

Claims

1. A purified thermostable enzyme derived from the eubacterium Thermosipho africanus, wherein said enzyme has a molecular weight of approximately 103,000 daltons and is characterized as having

(a) DNA polymerase activity;

(b) 5'"'"'→

3'"'"' exonuclease activity;

(c) 3'"'"'→

5'"'"' exonuclease activity; and

(d) a temperature optimum greater than 45°

C.

2. The enzyme of claim 1 consisting essentially of amino acids 1 through 892 of SEQ ID NO:

2.