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The DNKR mutant contains the following four mutations in the zinc-finger domain: D94A, N99A, K101A, and R102A.
Worms that did not respond to repeated stimulus were scored as dead and those that contained internally hatched larvae were excluded.
To determine whether the mutant contained the expected gene replacement, colony PCR was performed using F1 and R3 primers.
A mutant containing two nonadjacent mutations [14].
The double mutant contained both bands.
Rd LPS, from more sensitive "deep rough" mutants, contains only an inner saccharide core, whereas Ra LPS, from "rough" mutants, contains a longer polysaccharide region.
Notably, only mutants containing blades 2 4 were effectively secreted.
About 5% of the mutants containing transposition events were homozygous.
Srs5 mutants containing empty vectors were used as controls.
In addition to the wild-type cueO, we also identified 14 cueO mutants, including eight mutants containing the synonymous mutations and six mutants containing the non-synonymous mutations.
Mutant 3E7 contains eight amino acid substitutions.
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