Difference between revisions of "PGP1087"
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Revision as of 11:52, 27 March 2013
General comments
- Integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor, allows express of the downstream gene by the p(spac) promoter
- constructed in Jörg Stülke's lab (C. Diethmaier)
- E.coli: low copy plasmid, select for ampicillin
- B. subtilis: no ori, select for erythromycin/lincomycin
- based on pMUTIN-FLAG
Hints for the usage
- amplify the last 300bp of your gene of interest,
- skip the stop codon of the gene in the reverse primer
- take care that the integration is in-frame
Antibody
- dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)
The reference
