Difference between revisions of "PGP1870"
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'''General comments''' | '''General comments''' | ||
| − | Integrative [[plasmid]] for the fusion of | + | 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 | * ''E.coli'': multicopy plasmid, select for ampicillin | ||
* ''B. subtilis'': no ori, select for spectinomycin | * ''B. subtilis'': no ori, select for spectinomycin | ||
| Line 13: | Line 13: | ||
* 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 | ||
| − | ''' | + | '''The reference''' |
| − | + | <pubmed> 23475962 </pubmed> | |
| + | == Return to [[Plasmids]] == | ||
Latest revision as of 17:05, 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
- 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)
