Difference between revisions of "PGP172"

Latest revision as of 14:04, 3 June 2013

pGP172: click to enlarge

pGP172 cloning region: click to enlarge

vector for the expression of proteins in E. coli, the plasmid allows to fuse a Strep-tag to the N-terminus of the expressed protein

host: E. coli BL21(DE3)/pLysS, expression of the desired protein is induced by the addition of 1 mM IPTG to the culture

sequencing primer:
- CD13: 5’-AAACATATGGCTAGCTGGAGCCACCCGCAGTTC -3’
- NP20: 5’-GCAGCAGCCAACTCAGCTTCCTTTCGGGC-3’

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-vector for the expression of proteins in E. coli, the plasmid allows to fuse a Strep-tag to the N-terminus of the expressed protein
+* vector for the expression of proteins in ''E. coli'', the [[plasmid]] allows to fuse a Strep-tag to the N-terminus of the expressed protein
-host: E. coli BL21(DE3)/pLysS, expression of the desired protein is induced by the addition of 1 mM IPTG to the culture
+* host: ''E. coli'' BL21(DE3)/pLysS, expression of the desired protein is induced by the addition of 1 mM IPTG to the culture
-constructed in the [[Stülke]] lab
+* constructed in the [[Stülke]] lab
-in E. coli: ampicillin resistance
+* in ''E. coli'': ampicillin resistance
-based on pET3C
+* based on pET3C
-similar plasmid: [[pGP574]]
+* similar [[plasmid]]: [[pGP574]]
-sequencing primer:
+* sequencing primer:
-*'''CD13:''' 5’-AAACATATGGCTAGCTGGAGCCACCCGCAGTTC -3’
+**'''CD13:''' 5’-AAACATATGGCTAGCTGGAGCCACCCGCAGTTC -3’
-*'''NP20:''' 5’-GCAGCAGCCAACTCAGCTTCCTTTCGGGC-3’
+** '''NP20:''' 5’-GCAGCAGCCAACTCAGCTTCCTTTCGGGC-3’
+* important: ''Eco''RI, although mentioned in the MCS, is not unique!
-Merzbacher, M., Detsch, C., Hillen, W. & Stülke, J. (2004) ''Mycoplasma pneumoniae'' HPr kinase/ phosphorylase: Assigning functional roles to the P-loop and HPr kinase/ phosphorylase signature sequence motif. Eur. J. Biochem. 271: 367-374. [http://www.ncbi.nlm.nih.gov/sites/entrez/14717704 PubMed]
+<pubmed> 14717704 </pubmed>
+==  Return to [[Plasmids]] ==