Difference between revisions of "PGP1080"
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* take care that the integration is in-frame | * take care that the integration is in-frame | ||
* do not use the XmaIII site for cloning, as GFP is inserted via this restriction site | * do not use the XmaIII site for cloning, as GFP is inserted via this restriction site | ||
| + | == Return to [[Plasmids]] == | ||
Latest revision as of 13:57, 3 June 2013
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!!!
- 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
- do not use the XmaIII site for cloning, as GFP is inserted via this restriction site
