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In this instance, all genes associated to the metabolic reaction would have to be deleted to confirm essentiality of the reaction.
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The majority of the genes with a predicted and confirmed essentiality status were within the cholesterol metabolism.
26 Furthermore, nutrients known to be essential for healthy humans are also essential for patients, and therefore, the required documentation is not to confirm the essentiality of nutrients among patients, but rather to define when a certain form of nutritional support is more beneficial than leaving the patient to develop nutritional deficiencies.
To confirm the essentiality of OD, its expression was knocked down by RNAi.
Further study of each of these genes would be required to confirm their essentiality.
To confirm the essentiality of S158 for NobA activity, this residue was further mutated into an alanine.
To confirm the essentiality of the spr0479 gene, an ectopic copy was introduced into the spr1806 locus.
Although the fraction of technical failures was small, these observations reaffirm the need to confirm genetic essentiality by conditional methods, just as in conventional gene-by-gene studies.
A transposon mutant library of F. tularensis [14] further confirms the essentiality of the pathway, as MEP pathway knockouts are lethal, further validating the pathway as an attractive target in F. tularensis.
These results indicate that the second gene copy rescued cell viability in a Zn-inducible manner, and explicitly confirms the essentiality of spr0479.
Disruption of the pseudaminic acid biosynthesis genes pseB and pseC resulted in nonglycosylated flagellin, confirming their essentiality in the bacterial flagellum assembly.
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