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Since the replacement of hydrophilic glutamine with hydrophobic alanine may cause relatively large changes, we performed the Q453E mutation to eliminate the effect of the side chain rotation of Gln453.
In the case of UASsickle flies, we introduced a second azot mutation to eliminate azot function.
Alternatively, use of the same CD47 antibody with a D265A Heavy Chain mutation to eliminate FcR binding would be a useful contriol.
Because that study had utilized the Mstn loss-of-function mutation to eliminate myostatin function, we sought to determine whether the effects that we had observed in Dysf −/− mice were somehow specific to the F66 transgene by crossing the F66 transgene onto an mdx background.
Owing to the substantially low affinity of human CD32A/B, KD values were obtained for the bivalent Fc fusion versions of these receptors (whose Fc domain was inactivated via N297Q or D265A mutation to eliminate homologous interactions); this strategy provides adequate sensitivity, due to avidity effects, to determine binding differences.
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So how do we decide as a society which mutations to eliminate and which to promote?
FRET transfer rates between individual donor-acceptor pairs within these constructs were calculated from FRET efficiencies measured after systematically introducing point mutations to eliminate all other acceptors.
The A4V, L38V, G93A, L106V and S134N mutations were introduced into the AS-SOD1 background [21], denoted herein as WT, which contains the C6A/C111S mutations to eliminate irreversible unfolding reactions caused by disulfide interchange with the intra-subunit C57 C146 disulfide bond in the unfolded state or by spontaneous oxidation of the cysteines by molecular oxygen.
Transient knockdown can be combined with ENU or TILLING mutations to eliminate functional redundancy or perform epistasis experiments.
To rule out the possibility that this ubiquitous factor is playing a significant role in driving expression in these synthetic constructs, we repeated these experiments using a basal promoter carrying a mutation known to eliminate binding of this ubiquitous factor.
To control for possible pleiotropic effects of the mutation used to eliminate sperm (q785), sperm was introduced to fog-1 q785 fog-1 q785 glp-1 q224ts glp-1 q224tsth wildtype manimalsnd these animals were examined in parallel.
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