EC Number |
Protein Variants |
Reference |
---|
3.1.11.2 | D251A |
site-directed mutagenesis, the mutant shows a 3fold decline in the endonucleolytic cleavage activity |
-, 749954 |
3.1.11.2 | D277A |
mutant lacking all enzymatic activities |
682165 |
3.1.11.2 | E117D |
mutant is unable to cleave substrate in vitro without affecting substrate binding, substitution of Mn2+ for Mg2+ increases the activity |
650060 |
3.1.11.2 | E117Q |
mutant is unable to cleave substrate in vitro but can bind substrate, substitution of Mn2+ for Mg2+ does not increase the activity |
650060 |
3.1.11.2 | E39A |
inactive mutant enzyme ExoIII |
-, 752239 |
3.1.11.2 | E57A |
site-directed mutagenesis, the mutant shows a 2fold decline in the endonucleolytic cleavage activity |
-, 749954 |
3.1.11.2 | E57A/D251A |
site-directed mutagenesis, the double mutant shows no 3'-5' exonuclease activity |
-, 749954 |
3.1.11.2 | E58A |
altered Mg2+-binding properties, 3'-diesterase activity is reduced when compared to the native enzyme |
664070 |
3.1.11.2 | F242S |
site-directed mutagenesis, the mutant shows slightly increased 3'-5' exonuclease catalytic efficiency compared to wild-type |
749954 |
3.1.11.2 | more |
elucidation of the iVEC mechanism at the molecular level avances the development of in vivo DNA cloning technology. Multiple-fragment assembly of up to seven fragments in combination with an effortless transformation procedure using a modified host strain for iVEC is possible. In vitro recombination system for DNA cloning already allow the joining of multiple DNA fragments at once. Construction of an Escherichia coli strain that is optimized for in vivo cloning. Generation of exonuclease deletion mutants from strain BW25113, the parental strain of the Keio collection, has sufficient capacity for iVEC activity |
750978 |