Exonuclease III (E. coli)
- 3' → 5' exonuclease
- Production of unidirectional nested deletions
- Site-directed mutagenesis
- Preparation of strand-specific probes
- Preparation of single-stranded substrates for dideoxy sequencing
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Catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of duplex DNA (1). A limited number of nucleotides are removed during each binding event, resulting in coordinated progressive deletions within the population of DNA molecules (2).
The preferred substrates are blunt or recessed 3´-termini, although the enzyme also acts at nicks in duplex DNA to produce single-strand gaps. 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.
Exonuclease III activity depends partially on helical structure (4) 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 (1).
Product SourcePurified from E.coli K-12, BE257/pSGR3 strain (kindly supplied by B. Weiss)
The following reagents are supplied with this product:
Store at (°C) Concentration NEBuffer™ 1 -20 10 X
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