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Among these mutants, a double Lys to Arg mutant, i.e. K406R/K573R, showed the best thermostability improvement.
To test epistasis relationships between these mutants, a double mutant was constructed (see Methods).
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I then mutated one individual with one mutation, mutated a second individual with the second mutation, and mutated the third individual with both mutations, creating both single mutants and a double mutant.
Fig. 2A shows that unlike the tsc2Δ single mutant, a double null mutant of arn1+ and tsc2+ showed high sensitivity to canavanine as with arn1Δ.
To account for the loss of symmetry in a single Cys to Ser mutant, a double Cys to Ser construct was made.
To investigate the virulence of the double mutant, SCID mice were injected intravenously with 105 CFU of either wild-type M. tuberculosis (strain H37Rv), a ΔsecA2 mutant, or a double mutant strain ΔsecA2ΔlysA.
We observed that in contrast to wild-type or single set1Δ or clr3Δ mutant strains, a double mutant set1Δ clr3Δ strain exhibited a significant synthetic slow-growth phenotype and sensitivity to the tubulin inhibitor thiabendazole, suggesting defects in chromosome segregation.
A genetically interacting gene pair was identified by examining the growth curves of a single gene knockout mutant and a double gene knockout mutant.
The pbTybceABΔloop plasmid was also used to obtain the bceBΔloop strain from the bceAB mutant via a double recombination event.
The oxa1 mutant carries a double amino-acid substitution E65G-F229S in the Oxa1 protein [ 38].
The single mutant C320S and a double mutant with both cysteines replaced by serine have been compared with one another in terms of long-term stability and other properties.
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