Expression vector, expression system and application of recombinant TEV protease
A technology for expressing vectors and recombinant proteins, which is applied in the biological field and can solve problems such as low protein stability
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[0033] 1. Experimental materials, reagents and experimental instruments
[0034] 1.1 Experimental materials
[0035] E.coli BL21 (DE3) is a strain preserved in our laboratory.
[0036] The recombinant plasmid pET-21a-TEVp was donated by Dr. Michel Vincent, University of Laval, Canada. The schematic diagram of the structure of plasmid pET-21a is attached figure 1 shown.
[0037] The four-point mutant TEVp (T17S, L56V, S135G and S219V) gene was inserted into the multiple cloning site of the pET-21a plasmid, and the recombinant plasmid pET-21a-TEVp was transformed into E.coli BL21 (DE3) and screened positive by Amp resistance Cloning, IPTG induced expression to produce His-TEVp recombinant protein.
[0038] 1.2 Main reagents and sources
[0039] Ni 2+ -NTA-Sepharose is a product of Amersham Company. IPTG reagent is the product of Sigma Company. Yeast extract and peptone were from Oxiod. The plasmid extraction kit was from Tiangen Company. All other reagents obtained fr...
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