From SubtiWiki
Jump to: navigation, search
pGP1080: 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, allows express of the downstream gene by the P(spac) promoter

ATTENTION: This plasmid encodes the dimeric GFP that tends to produce artifacts!!!

  • 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
  • do not use the XmaIII site for cloning, as GFP is inserted via this restriction site

Return to Plasmids