We think that knocking out a specific gene in Neisseria meningitidis seems to increase recombination events (transformation efficiency, see graph 1).
In the overexpressed strain, the transformation efficiency is lowered, which supports this hypothesis (see graph 1).
To double-check that this phenotype is really due to the gene/protein I am studying, I recently made a complementation strain by adding the gene under a highly expressed promoter somewhere else in the genome. The complementation was proven successful by PCR and growth on selective media. However, the transformation efficiency is even higher than in the KO (see graph 2).
Can anybody explain that ?
ps : Don't mind the dKO (double-KO).
I have seen something similar happen with eukaryotic cells.
-The gene may need to be reintroduced with its native promoter/regulatory elements, and not in an overexpression plasmid.
It is possible to complement knockouts with recombinant protein.
-Your cells might have found a way to cope with the absence of the gene by upregulating/downregulating others.
-Your knockout might have unfortunately affected other genes.
All of the above are testable.
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