The procedure involved, first, optimizing the electrotransformation conditions of the Xac 306 strain and, second, constructing non-replicative suicide vectors carrying knockout copies of the target gene.
If experimental data on the knockout and copy-number mutants were available, their signals could be compared with these predictions and thus used to validate the model or to identify its weaknesses.
To investigate the effect of Hr6b knockout on single-copy and multi-copy X-linked genes, 552 single-copy and 25 multi-copy genes described by Mueller et al. [ 36], were selected in wild type and Hr6b knockout normalised microarray data of spermatocytes and spermatids.
Two types of mutants are particularly interesting: gene-knockout and copy-number mutants.
The CNV resulting from single knockout of one copy each of these two tandemly arrayed genes is obvious at the whole chromosome view of chromosome 35, panel A.
We first suspected that ssrA gene might be essential in S. coelicolor and thus incorporated additional controls into our knockout experiment including extra copy of wild type and mutant ssrA-DD.
Complementation of the knockout mutant with a copy of the gene on a plasmid transcribed from the ermE* promoter restored production to wild-type levels.
In contrast, knockout mice have neither copy of the gene nor the corresponding gene expression products, which may also explain the difference between human studies and mice experiments.
In littermates of the double knockout, when one functional copy of Myf5 is present (Myf5 nlacZ/+:Myod −/−), skeletal muscle differentiation occurs, but the number of muscle fibers, and muscle mass, is decreased by 35 55% [23].
Disruption of one gene leads to inhibition of cell growth, whereas knockout of both gene copies is lethal.
