I have been trying to express a kinase recombinant protein in BL21 DE3 and C43 but couldn't get any expression.
I used 2L LB culture and 0.5mM IPTG induction.
I have even tried with colonies from fresh transformation into fresh competent cells still couldn't observe expression.
I would like to know which steps could have been gone wrong?
Poor expression in bacteria could be the result of rare codon usage in the gene construct. Try using an expression strain that is engineered to express more of the tRNAs for rare codons, or have the gene synthesized with codon optimization for E. coli expression.
Here is a nice article from NEB that covers that and other subjetcs relevant to protein expression:
https://www.neb.com/tools-and-resources/feature-articles/bypassing-common-obstacles-in-protein-expression
an improve way to express any protein kinase in bacteria is to coexpress the lambda phosphatase see http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2741569/
it give good results from my own experience
hope his help
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