These results illustrate the large effects caused by small changes in structure on long-range conformational dynamics and ligand specificities within proteins, further supporting the importance of preserving wild-type dynamics in enzyme systems that rely on flexibility for function.
We introduce a new problem which arises in the course of our project to design virus variants to serve as potential vaccines: we wish to modify virus-length genomes to introduce large numbers of unique restriction enzyme recognition sites while preserving wild-type function by substitution of synonymous codons.
Internal tagging of Cse4 confers exclusive centromeric localization and preserves wild type phenotype.
Positions 88 and 94 maintained base pairing, but did not preserve the wild type bases.
Despite demonstrating increased ventricular fibrosis compared to the mdx mice), the Sgcg−/− D2 animal's cardiac function was preserved at wild type levels.
In order to further establish the comparative differences in the functional expression of Clc-1 in Mbnl mutant mice, we investigated whether the voltage-dependence of the peak (instantaneous) and steady-state I V plots of ICl were preserved in wild type, Mbnl1 ΔE3/ΔE3, Mbnl3 ΔE2 and Mbnl1 ΔE3/ΔE3/Mbnl3 ΔE2 animals.
To avoid interferon induction by U6 vectors, we recommend preserving the wild-type sequence around the transcription start site, in particular a C/G sequence at positions −1/+1, and we describe a simple cloning strategy using the Gateway recombination system that facilitates this task.
A series of mRNAs were prepared in which the original BM2fluc start codon was mutated to AGC and new AUG restart codons inserted independently at +12, +24, +36, +48 and +63 nt, again preserving the wild-type Kozak consensus.
All mutations introduced into the cloned cDNA fragments were translationally-silent, thereby preserving the wild-type polyprotein sequence.
CD spectra and X-ray crystallography showed that M118E-NTF2 preserved the wild-type NTF2 fold, although its thermal stability was 20 deg.
As reported previously, the majority of endogenous proteins labeled internally with a fluorescent tag preserve their wild-type localizations and functions [17], [18], [19].
