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Using sequence- and structure-based approaches, 48 CLF multiple mutants were engineered and analyzed.
By contrast, the multiple mutants were up to 8-fold less efficient than the recombinant wild-type dalcochinase, and displayed primarily antagonistic interactions between these residues.
The double mutants or multiple mutants were derived from genetic crosses of the parental mutants (or transgenic lines).
Multiple mutants were tested and only the variant present in the Pcsk5 gene was present in all of the affected mice.
In the last instance, the rpoB multiple mutants were selected in vitro in a stepwise fashion, and one double mutant, L511Q+D516G (also described in M. tuberculosis strain [ 3]), exhibited a relative fitness either greater than or equal to either single mutant or the wild type.
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In fact, competition environments in which multiple mutants are grown together can be rather different from medium containing only monocultures or two strains.
OX2 multiple point mutants were generated in the C-terminus using site-directed mutagenesis to replace serine and threonine residues with alanine.
Multiple-position mutants were not modeled.
We note however that the integrity of this loop appears to be critical for MCMJR1 activity, as multiple loop truncation mutants were found to be inactive in our in vitro assay (Fig. S1).
However, using multiple approaches, no mutants were isolated with unconditional disruptions in either Ccel_2137 (pta) or Ccel_2136 (ack) genes.
In addition to predictive power, MD simulations of multiple RT mutants are providing fundamental insight into the dynamics of the allosteric NNRTI binding site which is useful for the design of future inhibitors.
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