Difference between revisions of "PGP1080"

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(Created page with "250px|thumb|right|'''pGP1080: click to enlarge''' '''General comments''' * Integrative plasmid for the fusion of GFP tag to the C-terminus of a protei...")
 
 
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* 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
 
* 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
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== ATTENTION: This plasmid encodes the dimeric GFP that tends to produce artifacts!!! ==
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* constructed in [[Jörg Stülke]]'s lab (C. Diethmaier)
 
* constructed in [[Jörg Stülke]]'s lab (C. Diethmaier)
  
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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
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==  Return to [[Plasmids]] ==

Latest revision as of 13:57, 3 June 2013

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