Difference between revisions of "PBP43"
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* take care that the integration is in-frame | * take care that the integration is in-frame | ||
* do not use the HindIII site for cloning, as GFP is inserted via this restriction site | * do not use the HindIII site for cloning, as GFP is inserted via this restriction site | ||
| + | * Don't use XbaI and KpnI for cloning. They cut the vector twice! | ||
Revision as of 12:12, 1 March 2013
General comments
Integrative plasmid for the fusion of the monomeric variant of GFP A206K to the C-terminus of a protein, keeping the control of the expression under its natural promoter
- 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
- Don't use XbaI and KpnI for cloning. They cut the vector twice!
