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To study the nature of the contacts in detail, we used paramagnetic NMR relaxation enhancements, in combination with single-point mutations, to obtain distance constraints for the acid-unfolded ensemble of ACBP.
The simulation is run for a large number of 106 mutations to obtain long evolutionary trajectories which are used to calculate the averages.
As shown in Table VI, the combination of all eight mutations to obtain a HRP C1A variant without any N-glycans resulted in an enzyme variant with extremely reduced catalytic efficiency and thermal stability.
Because KCTD15 has two predicted binding sites in its 3′-UTR, we performed molecular cloning for single site mutations to obtain vectors named MUT1-KCTD15, MUT2-KCTD15, and double site mutation vectors named MUT1,2-KCTD15 MUT1,2-KCTD15
One of the most successful strategies has been to combine systematically a number of stabilizing mutations to obtain a modified protein that has been thermostabilised, while still retaining its native structure or at least a well-defined conformation that represents one of its natural conformations.
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These models formed the initial population on which the GA performed operations such as crossover and mutation to obtain model(s) with best rate constant prediction.
They start with a set of chromosomes called initial population and then go through (possibly) three operations, namely reproduction/selection, crossover, and mutation, to obtain a heuristically optimal solution.
Qu and He [47] presented an algorithm based on adaptive Gaussian mutation to obtain the global extremum and to improve the quality of optimal solutions as well as the efficiency of the algorithm.
The original stop codon of the luciferase gene was disrupted by a point mutation to obtain TCA-2E.
An advantage of using mutagens that result in large deletions is the possibility of detecting mutations knocking out tandem-duplicated gene family members – a difficult mutation to obtain by traditional mutagenesis methods.
In future studies, we may conditionally knockout the Loxl3 gene in specific tissues or knock-in the LOXL3 human mutation to obtain viable mice in which cleft palate or spinal deformity is still present.
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