PGP1870

From SubtiWiki
Revision as of 12:29, 3 November 2011 by MLehnik (talk | contribs)
Jump to: navigation, search
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

Antibody

  • dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)