USER (Uracil-Specific ExcisionReagent) Enzyme (1) generates a single nucleotidegap at the location of a uracil. USER Enzyme is amixture of Uracil DNA glycosylase (UDG) and theDNA glycosylase-lyase Endonuclease VIII. UDGcatalyses the excision of a uracil base, forming anabasic (apyrimidinic) site while leaving the phosphodiesterbackbone intact (2,3). The lyase activityof Endonuclease VIII breaks the phosphodiesterbackbone at the 3´ and 5´ sides of the abasic siteso that base-free deoxyribose is released (4,5).To help select the best DNA assembly method for your needs, please use our Synthetic Biology/DNA Assembly Selection Chart.Figure 2: USER Enzyme (NEB #M5505), Thermolabile USER II Enzyme (NEB #M5508) and Thermostable USER III Enzyme (NEB #M5509) generate different functional ends after cleavage of DNA. In addition to different optimal reaction temperatures (37°C for USER and Thermolabile USER II Enzyme and 65°C for Thermostable USER III Enzyme) and ability to be heat inactivated (Thermolabile USER II Enzyme only), the different USER Enzymes generate different 3’ and 5’ termini after cleavage. USER Enzyme (NEB #M5505) contains Endonuclease VIII and leaves a 3’ and 5’ phosphate after cleavage. Thermolabile USER II Enzyme (NEB #M5508) contains Endonuclease III and leaves a 3′-phospho-α, β-unsaturated aldehyde and 5′ phosphate after cleavage. Thermostable USER III Enzyme (NEB #M5509) contains Endonuclease IV and leaves a 3′-hydroxyl and 5′-deoxyribose phosphate.
Product Source
The two component proteins are purified separately from E. coli K-12 strains containing plasmids encoding Endonuclease VIII and Uracil-DNA Glycosylase.
This product is related to the following categories:
DNA Repair Enzymes and Structure-specific Endonucleases Products
This product can be used in the following applications:
Applications of USER® and Thermolabile USER II Enzymes,
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