Difference between revisions of "PGP1870"
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| − | 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 | ||
Revision as of 12:29, 3 November 2011
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
Antibody
- dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)
