The probes also showed their applicability to discriminate the 3′ → 5′ proofreading activity of different exonuclease-proficient DNA polymerases.
However, their participation in protein synthesis is generally prevented by an aaRS proofreading activity located in the synthetic site and in a separate editing domain.
Overall, our observations indicate that the exonuclease domain of the Klenow enzyme, besides its proofreading activity, does significantly contribute to the catalytic efficiency of DNA elongation.
DNA polymerases with intrinsic proofreading activity interact with DNA primer/templates in two distinct modes, corresponding to the complexes formed during the 5′ 3′ polymerization or 3′ 5′ editing of a nascent DNA chain.
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High fidelity enzymes particularly have 3′ 5′ proofreading activity that affects added dATP residue stability (Sambrook et al. 1978); the resulting PCR product will thus have blunt ends.
Phi29 DNA polymerase has been considered the optimal choice for MDA because it shows higher efficiency, higher fidelity and a lower error rate compared with Bst DNA polymerase which has no proofreading activity [10, 46].
Many DNA polymerases have a proofreading activity.
Similarly, the proofreading activity of Pol III prevents lesion bypass by translesion DNA polymerases if the translesion polymerase is not sufficiently processive [12].
Deficiencies in polymerase proofreading activity, for example, might cause an increase in mutation rates, whereas incorrect non-homologous end joining might result in deletions or other polymorphisms.
Although HIV-1 RT exhibits no exonucleolytic proofreading activity, it still retains a relatively high level of fidelity of DNA synthesis, i.e., about 2.5 6×10−4 base substitutions per site [2], [3].
