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Herein, a further increase in fluorescence in the new BFPvvD8 mutant, with three additional mutations from BFPvvD7, was made.
A C-cap mutant with three additional mutations introducing suitably spaced charged residues did not show formation of salt bridges, explaining why its stability was not increased further.
Consequently, SInv and 5 mutants including three mutants with single point substitution [Mut1 = P152 V, Mut2 = S85 V and Mut3 = K153F ], one mutant with two points [Mut4 = S305V-N463 V] and one mutant with three points [Mut5 = S85V-K153F-T271 V] were developed via site-directed mutagenesis and produced using Pichia pastoris as the host.
A mutant with three point substitutions, Lys175Met/Cys176Gly/Tyr273Leu, i.e., HaloTag, has a high apparent second-order rate constant, thus allowing the labeling reaction to reach completion even under low haloalkane ligand concentrations [260].
Additionally, a Tmod1 mutant with three point mutations in the LRR domain that resulted in disrupted localization in cardiac cells, was overexpressed.
Construction of the Translation-Stop Mutant Gene whip∗ The construction of a whip∗ mutant with three tandem stop codons was a two-steps process.
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This mutant with two cysteine to serine mutations, HPTP-BNΔ6-CΔ2-C90S-C109S, has good potentian as an optimal reporter.
To this end, we generated one quadruple-point mutant with four threonine residues mutated to alanine (A) (referred to as Ezh2-M 4A), and one triple-point mutant with a T372 residue remaining intact (referred to as Ezh2-M 3A372T).
The walls were crowded with framed images, as at the Barnes Foundation, except in this case the collection featured Edward Gorey illustrations, concept-art sketches of the demon from "Fantasia" ("I'm an obsessive Disney-villain guy"), and comic-book panels, including a Richard Corben drawing of a mutant with four breasts.
In the mutant with two symmetrical hydrogen bonds (FM197H) a single cooperative rupture of the two bonds was observed corresponding to an about twice stronger bond, rather than a sequential rupture of two individual bonds.
However, a mutant with two point mutations near the N terminus (Gln H6 replaced by Glu and Ala H10 (Kabat H9) replaced by Gly) crystallized in a form compatible with antigen-binding.
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