Exonuclease III (E. coli)
from
New England Biolabs
Description
• Isolated from a recombinant source
• 3'→5' exonucleIsolated from a recombinant source
• 3'→5' exonuclease
• Produces unidirectional nested deletions
• Site-directed mutagenesis
• Supplied with 10X Reaction Bufferase
• Produces unidirectional nested deletions
• Site-directed mutagenesis
• Supplied with 10X Reaction Buffer
Catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of duplex DNA. A limited number of nucleotides are removed during each binding event, resulting in coordinated progressive deletions within the population of DNA molecules.
The preferred substrates are blunt or recessed 3´-termini, although the enzyme also acts at nicks in duplex DNA to produce single-strand gaps. The enzyme is not active on single-stranded DNA, and thus 3´-protruding termini are resistant to cleavage. The degree of resistance depends on the length of the extension, with extensions 4 bases or longer being essentially resistant to cleavage. This property can be exploited to produce unidirectional deletions from a linear molecule with one resistant (3´-overhang) and one susceptible (blunt or 5´-overhang) terminus.
Exonuclease III activity depends partially on helical structure and displays sequence dependence (C>A=T>G; ref. 5). Temperature, salt concentration and the ratio of enzyme to DNA greatly affect enzyme activity, requiring reaction conditions to be tailored to specific applications. Exonuclease III has also been reported to have RNase H, 3´-phosphatase and AP-endonuclease activities.
Specifications/Features
Unit Definition:
One unit is defined as the amount of enzyme required to remove 200 base pairs from each end of linearized double-stranded phiX174 DNA (650 µg/ml) in 10 minutes at 30°C.