Hi all,
Earlier I got the recombinant protein expression in E. coli. Now, I repeat the expression from glycerol stock but i didnt get the protein expression.
I repeat the fresh clone and transformation even though not getting the expression,
I have checked any sequence mutation in the gene but there is no mutation.
I did expression with different IPTG concentrations and different temperatures like 15, 18, 25, 30 and 37 C.
In all conditions I didnt get the protein expression.
Please kindly help me to overcome this problem, how should I get the protein expression in E. coli?
Thanking you
If you are certain that you have used an IPTG-inducible promotor AND your protein isn't toxic, I'd say that you should check that your reagents are all fresh.
Did you sequence the promotor to make sure it's intact and that your insert isn't backwards?
Hello!You said earlier you got expression .Which condition was that?Which temperature?Did you repeat the same condition again?
Was your protein in soluble or pellet form?This situation happens when the protein goes into pellet form so protein expression is not reproducible.
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