Description: T4PolynucleotideKinase(PNK)catalyzesthetransferandexchangeoftheterminalgammapositionphosphateofATPtothe5'-hydroxylterminusofdouble-andsingle-strandedDNA,RNAandnucleoside3'-monophosphatemolecules(1).T4PNKalsoexhibits3'-phosphataseand2',3'cyclicphosphodiesteraseactivities.(2-6)
SpecificActivity:133,333U/mg
Application: -End-labelingDNAorRNAforprobesandDNAsequencing -Additionof5'-phosphatestooligonucleotidestoallowsubsequentligation -Removalof3'-phosphorylgroups
Source: PurifiedfromastrainofE.colithatexpressestherecombinantT4PolynucleotideKinasegene.
Suppliedin: 10mMTris-HCl 50mMKCl 0.1µMATP 1.0mMDTT 0.1mMEDTA 50%Glycerol pH7.4@25°C
Suppliedwith: 10xT4PolynucleotideKinaseBuffer
10xT4PolynucleotideKinaseBuffer: 700mMTris-HCl 100mMMgCl2 50mMDTT pH7.6@25°C
UnitDefinition: Oneunitisdefinedastheamountofenzymecatalyzingtheincorporationof1nmolof[32P](ATPdonor)in30minutesat37°Cin1xT4PolynucleotideKinaseReactionBuffer.
RecommendedStorageCondition:-20°C
Reference: 1.Richardson,C.C.(1981)P.D.Boyer(Eds.),TheEnzymes,14,pp.229-314.SanDiego:Academicpress. 2.Morse,D.P.etal.(1997)Biochemistry36,8429-8434. 3.Cameron,V.etal.(1977)Biochemistry16,5120-5126. 4.Wand,L.K.etal.(2002)Nucl.AcidsRes.30,1073-1080. 5.Galburt,E.,etal.(2002)Structure10,1249-1260. 6.Wang,L.K.,etal.(2002)EMBOJ.21,3873-3880. |