I have glycerol stocks of transformed competent cells containing p3xFLAG-CMV-10 with ampicillin resistance and my gene of interest. The gene is a mouse oligodenylate synthetase protein with a transmembrane region. I get very low yields every time I do miniprep (
Is there only a CMV promoter available on the plasmid? Or are there (probably even cryptic) promoter sites which work in bacteria? I would suspect expression of your protein which seems to be toxic for the cells.
Here is the vector map. Hope this helps!
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/Vector/1/p3xflag-cmv-9-10_expression_vector.pdf
p3xFLAG-CMV-10 has pBR322 origin of replication. pBR322 is a middle copy no. plasmid. You can refer miniprep manual fron Qiagen to get protocol how to increase yield.
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