Title: Methods for detection of methyl-CpG dinucleotides

Abstract

The invention provides methods for enriching methyl-CpG sequences from a DNA sample. The method makes use of conversion of cytosine residues to uracil under conditions in which methyl-cytosine residues are preserved. Additional methods of the invention enable to preservation of the context of me-CpG dinucleotides. The invention also provides a recombinant, full length and substantially pure McrA protein (rMcrA) for binding and isolation of DNA fragments containing the sequence 5'-C.sup.MeCpGG-3'. Methods for making and using the rMcrA protein, and derivatives thereof are provided.

Inventors:

Dunn, John J.

Issue Date:

Tue Jan 29 00:00:00 EST 2013

Research Org.:

Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Org.:

USDOE

OSTI Identifier:

1082684

Patent Number(s):

8361746

Application Number:

12/507,522

Assignee:

Brookhaven Science Associates, LLC (Upton, NY)

Patent Classifications (CPCs):

C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

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 {

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C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

C07K14/245 - Escherichia
C07K2319/40 - containing a tag for immunodetection, or an epitope for immunisation

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
C12P19/36 - Dinucleotides, e.g. nicotineamide-adenine dinucleotide phosphate

C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS

C12Q1/6827 - for detection of mutation or polymorphism
C12Q2522/101 - Single or double stranded nucleic acid binding proteins
C12Q2523/125 - Bisulfite

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DOE Contract Number:  

AC02-98CH10886

Resource Type:

Patent

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES