Compositions and Methods for the Transfer of a Hexosamine to a Modified Nucleotide in a Nucleic Acid

Abstract

Nucleic acids comprising β-glucosaminyloxy-5-methylcytosine; compositions, kits and methods of producing the nucleic acids using a glycosyltransferase; and methods of using the nucleic acids are described.

What is claimed is: 
     
         1 .- 18 . (canceled) 
     
     
         19 . A method, comprising:
 (a) attaching a hexosamine to a modified nucleotide in a nucleic acid fragment   (b) adding a label to the hexosamine to form a labeled modified nucleotide; and   (c) detecting the labeled modified nucleotide and/or enriching for the nucleic acid containing the labeled modified nucleotide.   
     
     
         20 . A method according to  claim 19 , wherein the hexosamine is glucosamine. 
     
     
         21 . A method according to  claim 19 , wherein the modified nucleotide is a hydroxymethylcytosine. 
     
     
         22 . A method according to  claim 19 , wherein the nucleic acid fragment is DNA. 
     
     
         23 . The method according to  claim 19 , wherein the nucleic acid fragment is a genomic fragment. 
     
     
         24 . The method according to  claim 19 , wherein (a) further comprises enzymatically attaching the hexosamine to the modified base. 
     
     
         25 . The method according to  claim 19 , further comprising covalently attaching the hexosamine to the modified base. 
     
     
         26 . The method according to  claim 19 , further comprising adding a label to the hexosamine by covalently linking the label to an amino group of the hexosamine containing nucleotides. 
     
     
         27 . The method according to  claim 19 , wherein the attached label is capable of binding to a solid support for separating the nucleic acid fragments with the modified nucleotide from nucleic acid fragments not containing the modified nucleotide. 
     
     
         28 . The method according to  claim 19 , wherein the attached label provides a signal capable of detection by a detector for detecting the modified nucleotide in the nucleic acid fragment. 
     
     
         29 . The method according to  claim 19 , wherein the attached label is capable of quenching a signal. 
     
     
         30 . The method according to  claim 19 , wherein the nucleic acid fragment is genomic DNA, the modified base is hydroxymethylcytosine, and the hexosamine is glucosamine. 
     
     
         31 . A kit comprising a purified β-glucosyl transferase and a purified UDP-glucosamine in a buffer in separate or the same reaction vessel. 
     
     
         32 . A nucleic acid comprising a modified nucleotide covalently attached to a hexosamine. 
     
     
         33 . The nucleic acid according to  claim 32 , wherein the modified nucleotide covalently attached to a hexosamine is a 5-(β-glucosaminyloxymethyl)cytosine. 
     
     
         34 . The nucleic acid according to  claim 33 , wherein the modified nucleotide covalently attached to a hexosamine is a 5-(β-2-glucosaminyloxymethyl)cytosine. 
     
     
         35 . The nucleic acid according to  claim 33 , wherein the modified nucleotide covalently attached to a hexosamine is a 5-(β-6-glucosaminyloxymethyl)cytosine.