NUCLEIC ACID BINDING PROTEINS AND USES THEREOF
Compositions, systems and methods employing nucleic acid binding proteins for use in the regulation and/or modulation of nucleic acid based reactions, including transcription, translation, modification, digestion, and hybridization reactions. Such compositions are employed in controlling a variety of different reaction types involving nucleic acids.
1 . A method of controlling a reaction with a nucleic acid, comprising:
providing the nucleic acid comprising at least one uracil containing nucleotide;
providing a nucleic acid binding protein with specificity for binding uracil containing nucleic acids in contact with the nucleic acid, under conditions whereby the nucleic acid binding protein binds to a portion of the nucleic acid comprising the uracil containing nucleotide, whereby binding of the nucleic acid protein to the nucleic acid at least partially blocks the reaction with the nucleic acid.
2 . The method of claim 1 , wherein the reaction comprises a ligation reaction, a polymerization reaction, an exonuclease reaction, an endonuclease reaction, a protection reaction, and/or a hybridization reaction.
3 . The method of claim 2 , wherein the reaction comprises a polymerization reaction.
4 . The method of claim 1 , wherein the nucleic acid binding protein comprises an archeal polymerase, a uracil-DNA glycosylase, a uracil binding fragment or a construct thereof.
5 . The method of claim 4 , wherein the nucleic acid binding protein comprises an archeal polymerase, a uracil binding fragment or a construct thereof.
6 . The method of claim 2 , wherein the reaction comprises a hybridization reaction and a reagent in the reaction comprises a polynucleotide complementary to at least a portion of the nucleic acid bound by the nucleic acid binding protein.
7 . The method of claim 2 , wherein the reaction comprises a protection reaction, and a reagent in the reaction comprises an exonuclease or an endonuclease, and the nucleic acid binding protein reduces activity of such endonuclease or exonuclease on at least a portion of the nucleic acid bound by the nucleic acid binding protein.
8 . A composition, comprising:
a nucleic acid comprising one or more uracil containing bases;
a nucleic acid binding protein with specificity for binding uracil containing nucleic acids, bound to the nucleic acid; and
a reagent capable of reacting with the nucleic acid, wherein presence of a bound nucleic acid binding protein at least partially blocks the reagent from reacting with the nucleic acid.
9 . The composition of claim 8 , wherein the reagent comprises an enzyme selected from the group consisting of a polymerase, a ligase, a transcriptase, an endonuclease and an exonuclease.
10 . The composition of claim 9 , wherein the reagent comprises a polymerase.
11 . The composition of claim 8 , wherein the nucleic acid binding protein comprises an archeal polymerase, a uracil-DNA-glycosylase, a uracil binding fragment or a construct thereof.
12 . The composition of claim 11 , wherein the nucleic acid binding protein comprises an archeal polymerase, a uracil binding fragment or a construct thereof.
13 . The composition of claim 12 , wherein the archeal polymerase comprises 9° North polymerase, a uracil binding fragment or a construct thereof.
14 . The composition of claim 13 , wherein the 9° North polymerase comprises a polymerase inactive construct of 9° North polymerase.
15 . The composition of claim 13 , wherein the 9° North polymerase comprises an exonuclease deficient 9° North polymerase.