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 Streptag tag, keeping the control of expression under control of the natural promotor of the target gene.
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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

pGP1389: click to enlarge

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)