Difference between revisions of "PGP1389"
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* skip the Stopp-Codon of the gene in the reverse primer | * skip the Stopp-Codon of the gene in the reverse primer | ||
* take care that the integration is in-frame | * take care that the integration is in-frame | ||
| + | |||
| + | '''The reference''' | ||
| + | <pubmed>21803996</pubmed> | ||
Revision as of 12:54, 16 August 2011
General comments
Integrative plasmid for the fusion of the C-terminus of a protein to a Streptag 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)
