PGP1087

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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 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

Christine Diethmaier, Nico Pietack, Katrin Gunka, Christoph Wrede, Martin Lehnik-Habrink, Christina Herzberg, Sebastian Hübner, Jörg Stülke
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] [WorldCat.org] [DOI] (I p)

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