I am performing a protein localization experiment on E.coli using a complimented strain which has a pBAD vector. I need to get this plasmid based expression to native protein level. Also I am hoping to conduct an immune-fluorescence experiment to observe it.
one could try to detect the target protein in WT (by activity or WB) and tune
the arabinose to match the WT expression levels by expressing the protein from
plasmid.
Instead of a plasmid driven expression, one can consider epitope tagging of the genomic copy of your favorite gene in order to get expression of the tagged protein under the control of the natural promoter of the gene (Northern blot should be used to compare RNA levels in WT and constructed strain). If the gene is essential, survival of the constructed strain will indicate the tagged protein is functional
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