Difference between revisions of "PGP1389"
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'''General comments''' | '''General comments''' | ||
| − | Integrative [[plasmid]] for the fusion of the C-terminus of a protein to a | + | Integrative [[plasmid]] for the fusion of the C-terminus of a protein to a Strep tag, keeping the control of expression under control of the natural promotor of the target gene. |
* ''E.coli'': multicopy plasmid, select for ampicillin | * ''E.coli'': multicopy plasmid, select for ampicillin | ||
Revision as of 11:33, 30 October 2012
General comments
Integrative plasmid for the fusion of the C-terminus of a protein to a Strep tag, keeping the control of expression under control of the natural promotor of the target gene.
- E.coli: multicopy plasmid, select for ampicillin
- B. subtilis: no ori, select for spectinomycin
Hints for the utilisation of the plasmid
- 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
The reference
Martin Lehnik-Habrink, Joseph Newman, Fabian M Rothe, Alexandra S Solovyova, Cecilia Rodrigues, Christina Herzberg, Fabian M Commichau, Richard J Lewis, Jörg Stülke
RNase Y in Bacillus subtilis: a Natively disordered protein that is the functional equivalent of RNase E from Escherichia coli.
J Bacteriol: 2011, 193(19);5431-41
[PubMed:21803996]
[WorldCat.org]
[DOI]
(I p)
