Difference between revisions of "PGP1389"
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'''General comments''' | '''General comments''' | ||
| − | Integrative plasmid for the fusion of | + | 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 | * ''E.coli'': multicopy plasmid, select for ampicillin | ||
* ''B. subtilis'': no ori, select for spectinomycin | * ''B. subtilis'': no ori, select for spectinomycin | ||
| − | '''Hints for the | + | '''Hints for the utilisation of the plasmid''' |
* amplify the last 600bp of your gene of interest, | * amplify the last 600bp of your gene of interest, | ||
* 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 | ||
Revision as of 19:05, 18 October 2010
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
