Title: Restriction/modification polypeptides, polynucleotides, and methods

Abstract

The present invention relates to the discovery of a novel restriction/modification system in Caldicellulosiruptor bescii. The discovered restriction enzyme is a HaeIII-like restriction enzyme that possesses a thermophilic activity profile. The restriction/modification system also includes a methyltransferase, M.CbeI, that methylates at least one cytosine residue in the CbeI recognition sequence to m.sup.4C. Thus, the invention provides, in various aspects, isolated CbeI or M.CbeI polypeptides, or biologically active fragments thereof; isolated polynucleotides that encode the CbeI or M.CbeI polypeptides or biologically active fragments thereof, including expression vectors that include such polynucleotide sequences; methods of digesting DNA using a CbeI polypeptide; methods of treating a DNA molecule using a M.CbeI polypeptide; and methods of transforming a Caldicellulosiruptor cell.

Inventors:

Westpheling, Janet; Chung, DaeHwan; Huddleston, Jennifer; Farkas, Joel A

Issue Date:

2015-02-24

Research Org.:

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

Sponsoring Org.:

USDOE

OSTI Identifier:

1171089

Patent Number(s):

8962333

Application Number:

13/439,069

Assignee:

University of Georgia Research Foundation, Inc. (Athens, GA)

Patent Classifications (CPCs):

C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

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C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES

C12Y301/21004 - Type II site-specific deoxyribonuclease

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N9/1007 - {Methyltransferases

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

C12P19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides

C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES

C12Y201/01037 - DNA

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N9/22 - Ribonucleases {RNAses, DNAses

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DOE Contract Number:  

AC05-00OR22725

Resource Type:

Patent

Resource Relation:

Patent File Date: 2012 Apr 04

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES