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URL: http://www.addgene.org/8835
Proper Citation: RRID:Addgene_8835
Insert Name: tobacco etch virus protease, catalytic domain
Organism: tobacco etch virus
Bacterial Resistance: Chloramphenicol and Ampicillin
Defining Citation: PMID:11809930
Vector Backbone Description: Backbone Marker:New England Biolabs; Backbone Size:6700; Vector Backbone:pMal-C2; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol and Ampicillin
Comments: pRK1043 overproduces the catalytic domain of tobaco etch virus (TEV) protease in the form of an affinity sandwich, with E. coli maltose-binding protein fused to its N-terminus and a polyarginine tag fused to its C-terminus. TheTEV protease produced by this strain is the autoinactivation-resistant mutant S219V.The pRIL plasmid (Stratagene) improves the yield of TEV protease by increasing the supply of argU tRNA in the cell (for AGG and AGA codons). The catalytic activity of the S219V mutant is approximately 2-fold greater than that of the wild-type protease. Shifting the temperature from 37°C to 30°C during induction maximizes the yield of soluble MBP-TEV-Arg fusion protein. Bacterial strain: E.coli BL21(DE3)-RIL (Stratagene).
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Source: Addgene
