Hello. I am working with a sodC mutant (JW1638) and according to Coli Genetic Stock Center, it's derived from K-12. Here is the genotype:
F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, ΔsodC724::kan, rph-1, Δ(rhaD-rhaB)568, hsdR514
I would like to do some complementation assays with this mutant and I would like to know which promoters does this strain recognize. For exemple, if I use a vector with a lac promoter, can I induce the expression of my gene in this mutant with IPTG?
The lac promoter on your vector will still function, however unless you are supplying the lacI repressor from your plasmid, there will not be sufficient repressor to regulate a multi copy plasmid so the expression of your gene is likely to be mostly constitutive (unregulated).
Hello Daniel,
I think the short answer to your question is yes. The ΔlacZ4787(::rrnB-3) portion of your strain's genome means that the terminator sequence from the 16S gene has been added (in triplicate) to the N-terminal portion of the lacZ gene basically generating a lacZ deletion. So technically the promoter still exists. However, if you introduce a vector with the lac promoter and induce with IPTG you will get expression off of your vector. Expression from the genome will technically happen but will produce a very short lacZ (probably will never see it).
The arabinose operon (araBAD) is deleted from your strain (from about 25 bp upstream of araB to 8 bp into araD) so that promoter is likely not recognized by the strain. So if you use a pBAD expression vector that should work too.
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