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help needed with prokaryotic expression
I tried to express a segment about 600 bp inserted in pET30 in BL21 strain. The ORF is correct by sequencing, however, the protein is not expressed at 30 or 37 degree with 0.1 to 1 mM IPTG induction.
I appreciate very much for any suggestions.
Re: help needed with prokaryotic expression
How do you determine whether or not your PoI is expressed?
I didnot find the pET30 vector; I do find the pET-30a/b/c on Addgene. What I see, is the ORF regulated by Lac operon is in the anti-clockwise direction. Are you sure that your insert is not reverted?
best and good luck
|expression , needed , prokaryotic|
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