PGP1870

pGP1870: click to enlarge

General comments

Integrative plasmid for the fusion of GFP tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor

• E.coli: multicopy plasmid, select for ampicillin
• B. subtilis: no ori, select for spectinomycin

Hints for the usage

• amplify the last 600bp of your gene of interest,
• skip the Stopp-Codon of the gene in the reverse primer
• take care that the integration is in-frame
• do not use the HindIII site for cloning, as GFP is inserted via this restriction site

The reference

Fabian M Rothe, Christoph Wrede, Martin Lehnik-Habrink, Boris Görke, Jörg Stülke
Dynamic localization of a transcription factor in Bacillus subtilis: the LicT antiterminator relocalizes in response to inducer availability.
J Bacteriol: 2013, 195(10);2146-54
[PubMed:23475962] [WorldCat.org] [DOI] (I p)