Modified DNA-polymerase from carboxydothermus hydrogenoformans and its use for coupled reverse transcription and polymerase chain reaction

• US 6,794,177 B2
• Filed: 02/05/2002
• Issued: 09/21/2004
• Est. Priority Date: 12/02/1997
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Original Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Inventors: Achhammer, Gunthar, Ebenbichler, Christine, Ankenbauer, Waltraud, Markau, Ursula
• Primary Examiner(s): HUTSON, RICHARD G
• Application Number:
  US10/072,436
• Publication Number:
  US 20020160399A1
• Time in Patent Office: 959 Days
• Field of Search: 435/23.7, 435/194, 435/183, 435/320.1, 435/252.3, 530/350, 536/23.5, 536/23.1, 536/23.2
• US Class Current: 435/252.3
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Abstract

A purified DNA polymerase exhibiting reverse transcriptase activity in the presence of magnesium ions and/or manganese ions having reduced or no 5′-3′-exonuclease activity and substantially no RNaseH activity and obtainable from Carboxydothermus hydrogenoformans.

Claims

1. An isolated DNA sequence encoding SEQ ID NO:

11.

2. The DNA sequence of claim 1 that comprises SEQ ID NO:

10.

3. A vector comprising the DNA sequence of claim 1.

4. A vector comprising the DNA sequence of claim 2.

5. The vector of claim 4 that is pΔ

_2-225AR₄.

6. A host cell transformed with the vector of claim 3.

7. A host cell transformed with the vector of claim 4.

8. A host cell transformed with the vector of claim 5.

9. The host cell of claim 8 that is E. coli GA1.