# US 5,700,672 A · Purified thermostable pyrococcus furiousus DNA ligase

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- Number: US 5,700,672 A
- Document ID: US5700672A
- Status: Granted
- Kind: A
- Application number: 07/919,140
- Filed: 1992-07-23
- Granted: 1997-12-23
- Earliest claimed: 1992-07-23
- Assignees: Stratagene
- Inventors: Eric J. Mathur; Edward J. Marsh; Warren E. Schoettlin
- Web page: https://patentgrep.com/patent/US5700672A

## CPC

- C12N 9/93: {Ligases (6)}
- Y10S 435/822: using bacteria or actinomycetales
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Purified thermostable DNA ligase is described that catalyzes template-dependent ligation at temperatures of about 30.degree. C. to about 80.degree. C., and which substantially retains its catalytic ability when subjected to temperatures of from about 85.degree. C. to about 100.degree. C. The thermostable DNA ligase has an estimated molecular weight of 50,000 to 70,000 daltons. A preferred thermostable DNA ligase is described that was isolated from the archaebacteria Pyrococcus furiosus. Also described are plasmid vectors for producing recombinant thermostable DNA ligase.

## Claims

### Claim 1

A purified thermostable DNA ligase from Pyrococcus furiosus having the amino acid sequence of SEQ ID NO: 1 which substantially retains activity when subjected to temperatures of from about 85.degree. C. to about 100.degree. C.

### Claim 2 (depends on 1)

The ligase of claim 1 that is isolated from a recombinant organism transformed with a vector that codes for the expression of said DNA ligase.

### Claim 3 (depends on 1)

A ligase according to claim 1, wherein the ligase is encoded by plasmid pEM1 (ATCC Deposit No. 75259).