# US 2014/0193877 A1 · Thermus brockianus nucleic acid polymerases

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- Number: US 2014/0193877 A1
- Document ID: US20140193877A1
- Status: Published application
- Kind: A1
- Application number: 14/106,183
- Filed: 2013-12-13
- Published: 2014-07-10
- Earliest claimed: 2001-11-30
- Assignees: Applied Biosystems, LLC
- Applicants: Applied Biosystems, LLC
- Inventors: Elena BOLCHAKOVA; James ROZZELLE
- Web page: https://patentgrep.com/patent/US20140193877A1

## CPC

- C12N 9/1252: {DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase}
- C12N 9/1241: {Nucleotidyltransferases (2.7.7)}
- C12P 19/34: Polynucleotides, e.g. nucleic acids, oligoribonucleotides
- C12Q 1/6869: Methods for sequencing
- C12Y 207/07007: DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
- Y02P 20/52: using catalysts, e.g. selective catalysts
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Y: ENZYMES
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The invention provides nucleic acids and polypeptides for nucleic acid polymerases from a thermophilic organism, Thermus brockianus. The invention also provides methods for using these nucleic acids and polypeptides.

## Claims

### Claim 1

An isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.

### Claim 2 (depends on 1)

The isolated nucleic acid of claim 1, wherein said nucleic acid polymerase comprises a mutation that decreases 5-3′ exonuclease activity.

### Claim 3 (depends on 2)

The isolated nucleic acid of claim 2, wherein said decreased 5-3′ exonuclease activity is relative to a nucleic acid polymerase that does not comprise said mutation.

### Claim 4 (depends on 1)

The isolated nucleic acid of claim 1, wherein said nucleic acid polymerase comprises a mutation that reduces discrimination against dideoxynucleotide triphosphates.

### Claim 5 (depends on 4)

The isolated nucleic acid of claim 4, wherein said reduced discrimination against dideoxynucleotide triphosphates is relative to a nucleic acid polymerase that does not comprise said mutation.

### Claim 6

An isolated nucleic acid comprising the nucleotide sequence of any one of SEQ ID NO:1-8 or a nucleotide sequence complementary to any one of SEQ ID NO:1-8.

### Claim 7

(canceled)

### Claim 8

An isolated nucleic acid encoding a nucleic acid polymerase from Thermus brockianus comprising amino acid sequence with at least 96% identity to any one of SEQ ID NO:9-16.

### Claim 9 (depends on 8)

A vector comprising the isolated nucleic acid of claim 8.

### Claim at position 10

-16. (canceled)

### Claim 17 (depends on 8)

An expression vector comprising a promoter operably linked to the isolated nucleic acid of claim 8.

### Claim at position 12

-24. (canceled)

### Claim 25 (depends on 8)

A host cell comprising the isolated nucleic acid of claim 8.

### Claim at position 14

-54. (canceled)