Description
Description
Expression plasmid. The expression cassette is under transcriptional control of the tetracycline promoter/operator. The expressed recombinant protein will be secreted into the periplasm Affinity tagStrep-Tactin® affinity tag (Strep-tag II®) for the purification of recombinant protein. The affinity tag is fused to the N-terminus of the recombinant protein and can be removed by cleavage with factor Xa.
Secretion The ompA signal sequence directs the expressed protein into the periplasmic space and will be cleaved off during the translocation process. Bacterial Expression Expression is induced upon addition of 200 ug anhydrotetracycline (order no.: 2 0401 001; 2 0401 002) per 1 liter E. coli shaking culture (A550= 0.5). Expression strainAny E. coli strain. The tet promoter works independently from the genetic background of E. coli.
Resistance
Chloramphenicol. Note: The CamR resistance gene codes for homotetrameric chloramphenicol acetyltransferase (MW of the monomer = 26.6 kDa) which is predominantly expressed in the cytosol of E.coli transformed with this plasmid.
Form 5 ug, dissolved in 10 mM Tris/HCl pH 8.0, 1 mM EDTA; 20 ul. Concentration 250 ng/ul. Storage 4 °C for frequent usage, 20 °C for long term storage.
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For research use only. Not for diagnostic or clinical use.