The mutated construct, pAChE-M-Luc, derived from pAChE-Luc, was constructed by site-directed mutagenesis, which we accomplished using mutagenic primers and flanking primers.
Beyond that distance, there is no significant difference (p = 0.0032 at 2 kb and 0.00006 at 8 kb, and 0.28, 0.1, 0.074, 0.075, 0.051, 0.362, and 0.16 for the remaining distances, respectively; Figure 3B), implying that in both WT and recD, low-level long-distance mutagenesis, which we propose results from processive BIR, is functional.
We used the same active-site targeted saturation mutagenesis libraries from which we previously identified mutants with improved activity towards glycolaldehyde, which is C2-hydroxylated like all natural TK substrates.
As an alternative strategy, we therefore used insertional mutagenesis, which can rapidly lead to the cloning of the relevant gene.
Here, we will describe only mutagenesis, which has partially been improved from the previous version of TRITON, and docking, which is a new option not included in the previous version of TRITON.
Combined with residue spatial fluctuations as determined from MD simulation, hydrogen bond network analysis and homologue analysis, we proposed the mutagenesis of selected residues which we hypothesized to be involved in protein-protein interaction.
Summing up, in this work we present another mechanism, SSR-localized mutagenesis, which might act together with inducible and stable hypermutability enhancing mutagenesis in mucA.
If this is the case, then how can we understand the concept of lethal mutagenesis, which has recently proven successful in a variety of viruses [ 59- 62], and for which the error threshold is generally cited as the underlying theory?
Using ClosTron and allelic exchange mutagenesis, which was demonstrated for the first time in an acetogen, we generated single mutants as well as double mutants for both aor and adhE isoforms to confirm the role of each gene.
In this study, we attempted to produce mutant strains through chemical mutagenesis, which could produce curdlan with improvement in yield and productivity than the wild strain.
We further identified positively charged surface residues by KS mutagenesis, which mediates key interactions with the negatively charged ACP surface.
