Difference between revisions of "PGP1871"
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'''General comments''' | '''General comments''' | ||
| − | Integrative [[plasmid]] for the fusion of | + | 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 | ||
Latest revision as of 13:23, 2 April 2015
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)
