Difference between revisions of "PGP1871"

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'''General comments'''
 
'''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
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Integrative [[plasmid]] for the fusion of YFP 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
 
* ''E.coli'': multicopy plasmid, select for ampicillin
 
* ''B. subtilis'': no ori, select for spectinomycin
 
* ''B. subtilis'': no ori, select for spectinomycin
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* do not use the HindIII site for cloning, as GFP is inserted via this restriction site
 
* do not use the HindIII site for cloning, as GFP is inserted via this restriction site
  
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'''The reference'''
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<pubmed> 23475962 </pubmed>
 
==  Return to [[Plasmids]] ==
 
==  Return to [[Plasmids]] ==

Latest revision as of 13:23, 2 April 2015

pGP1871: click to enlarge

General comments

Integrative plasmid for the fusion of YFP 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)

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