FEN-1 endonuclease, mixtures and cleavage methods

• US 6,555,357 B1
• Filed: 10/06/2000
• Issued: 04/29/2003
• Est. Priority Date: 11/29/1996
• Status: Expired due to Term

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Third Wave Technologies Incorporated (Hologic Incorporated)
• Inventors: Lyamicheva, Natasha, Lyamichev, Victor I., Kaiser, Michael W.
• Primary Examiner(s): Patterson, Jr., Charles L.
• Application Number:
  US09/684,938
• Time in Patent Office: 935 Days
• Field of Search: 435/320.1, 435/252.9, 435/825, 435/194, 536/23.2
• US Class Current: 435/252.3
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 2525/301 Hairpin oligonucleotides
• CPC Class Codes: C12Q 2561/109 Invader technology
• CPC Class Codes: Y10S 435/975 Kit

Abstract

The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to improved cleavage means for the detection and characterization of nucleic acid sequences. Structure-specific nucleases derived from a variety of thermostable organisms are provided. These structure-specific nucleases are used to cleave target-dependent cleavage structures, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

Claims

1. A composition comprising an isolated oligonucleotide encoding a thermostable Archaeoglobus fulgidus FEN-1 endonuclease.

2. A composition comprising a recombinant DNA vector comprising the isolated oligonucleotide of claim 1.

3. A host cell comprising the recombinant DNA vector of claim 2.

4. The composition of claim 1, wherein said oligonucleotide comprises SEQ ID NO:

178.

5. A kit comprising the isolated oligonucleotide of claim 1.

6. A kit comprising two oligonucleotide primers, at least one of said two oligonucleotide primers selected from the group consisting of SEQ ID NOs:

170 and 171.

7. The kit of claim 6, further comprising a polymerase.

8. The kit of claim 6, further comprising a nucleic acid encoding a FEN-1 endonuclease.