Bacterial Strains Gold DE3 - (May/10/2006 )

I am trying to take expression of my proteins in BL21 Gold DE3 and Codon Plus strains.
In manual its written that these strains are also resistant to tetracyline. I am at the moment using amilicine for growing my transfromaton and expression culture. I am not getting expression and It take time to get tetracyline from the supplier. I want to ask that is it work with just ampiciline or u have to add both the antibiotic at the same time in single medium. I mean I have to use just amicicline or tetracyline alone or I have to add both antibiotics at the same time togather in single culture. I am using bacterial strains BL21 GOLD DE3 or BL21 GOLD CODON +.
best regards

All you need is ampicillin.

If you grow an overnight culture with 100 ug/ml ampicillin, do you see growth?

If you do and you still obtain no expression after induction with 0.2 mM IPTG overnight, then there is something wrong with your construct or your protein simply does not express that well.

May I ask what you are trying to express? Soluble, membrane protein, eukaryotic, prokaryotic origin, etc.?

-Matt

I am trying to express membrane protein vanilloid receptor TRPV1 thats is eukayotic origin and I did not get expression at all.