PGP1087

pGP1087: click to enlarge

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 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 references

Diethmaier C, Pietack N, Gunka K, Wrede C, Lehnik-Habrink M, Herzberg C, Hübner S, Stülke J    
A Novel Factor Controlling Bistability in Bacillus subtilis: The YmdB Protein Affects
Flagellin Expression and Biofilm Formation. 
J Bacteriol.: 2011, 193(21):5997-6007. 
PubMed:21856853