Type II restriction modification system methylation subunit of Alicyclobacillus acidocaldarius

Patent metadata

Field	Value
Publication number	US-9567595-B2
Application number	US-201514707231-A
Country	US
Kind code	B2
Filing date	May 8, 2015
Priority date	May 5, 2010
Publication date	Feb 14, 2017
Grant date	Feb 14, 2017

Abstract

Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods for modulating or altering recombination inside or outside of a cell using isolated and/or purified polypeptides and/or nucleic acid sequences from Alicyclobacillus acidocaldarius.

First claim

What is claimed is:

1. A vector comprising a first nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to SEQ ID No: 2891 and a second nucleic acid sequence heterologous to the first nucleic acid sequence.

2. The vector of claim 1 , wherein the polypeptide has activity at or below about pH 8.

3. The vector of claim 1 , wherein the polypeptide has activity at a temperature at or above about 50 degrees Celsius.

4. The vector of claim 1 , wherein the polypeptide has activity as a DNA adenine methylase.

5. A vector comprising a first nucleic acid sequence encoding the polypeptide of SEQ ID NO: 2891 and a second nucleic acid sequence heterologous to the first nucleic acid sequence.

6. The vector of claim 5 , wherein the first nucleic acid sequence comprises SEQ ID NO: 2892.

7. A method of modulating or altering nucleic acid methylation in a cell, the method comprising:
providing the vector of claim 1 to a cell; and
expressing the first nucleic acid sequence to produce the polypeptide.

8. The method according to claim 7 , wherein the modulation or alteration of nucleic acid methylation occurs at or below about pH 8.

9. The method according to claim 7 , wherein the modulation or alteration of nucleic acid methylation occurs at a temperature at or above about 50 degrees Celsius.

10. The method according to claim 7 , wherein the polypeptide is glycosylated, pegylated, or otherwise post-translationally modified.

11. The method according to claim 7 , wherein the polypeptide has activity as a DNA adenine methylase.

12. The method according to claim 7 , wherein the polypeptide has activity at or below about pH 8.

13. The method according to claim 7 , wherein the polypeptide has activity at a temperature at or above about 50 degrees Celsius.

14. A method of modulating or altering nucleic acid methylation, the method comprising:
providing an isolated or purified polypeptide having at least 90% sequence identity to SEQ ID No: 2891 to a nucleotide sequence in which adenine demethylation is desired.

15. The method according to claim 14 , wherein the modulation or alteration of nucleic acid methylation occurs at or below about pH 8.

16. The method according to claim 14 , wherein the modulation or alteration of nucleic acid methylation occurs at a temperature at or above about 50 degrees Celsius.

17. The method according to claim 14 , wherein the purified or isolated polypeptide is glycosylated, pegylated, or otherwise post-translationally modified.

18. The method according to claim 14 , wherein the purified or isolated polypeptide has activity as a DNA adenine methylase.

19. The method according to claim 14 , wherein the purified or isolated polypeptide has activity at or below about pH 8.

20. The method according to claim 14 , wherein the purified or isolated polypeptide has activity at a temperature at or above about 50 degrees Celsius.

21. The vector of claim 5 , wherein the polypeptide has activity at or below about pH 8.

22. The vector of claim 5 , wherein the polypeptide has activity at a temperature at or above about 50 degrees Celsius.

23. The vector of claim 5 , wherein the polypeptide has activity as a DNA adenine methylase.

24. A vector comprising a first nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to SEQ ID No: 2891 and a second nucleic acid sequence heterologous to the first nucleic acid sequence.

25. The vector of claim 24 , wherein the polypeptide has activity at or below about pH 8.

26. The vector of claim 24 , wherein the polypeptide has activity at a temperature at or above about 50 degrees Celsius.

27. The vector of claim 24 , wherein the polypeptide has activity as a DNA adenine methylase.

Assignees

• Battelle Energy Alliance Llc

Inventors

• Lee Brady D
• Newby Deborah T
• Lacey Jeffrey A
• Thompson David N
• Thompson Vicki S
• Apel William A
• Roberto Francisco F
• Reed David W

Classifications

• C12N9/00 Primary

  Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; {enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113}); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt C12C1/00) · CPC title

• C12N9/52

  derived from bacteria {or Archaea} · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12N9/1007

  Methyltransferases (general) (2.1.1.) · CPC title

• C12Y201/01072

  Site-specific DNA-methyltransferase (adenine-specific) (2.1.1.72) · CPC title