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Proteins, Expression, Isolation and Analysis> | … Dialysis bags were purchased from Union Carbide (Shanghai, China). An endotoxin assay kit was purchased from Genscript (Nanjing, China). 2.2. Optimized expression of Trx-BjαIT. BjαIT cDNA was amplified by polymerase chain reaction according to previous reports [16] … | Get A Quote |
Scorpion long-chain insect neurotoxins have important potential application value in agricultural pest control. The difficulty of obtaining natural toxins is the major obstacle preventing analyses of their insecticidal activity against more agricultural insect pests. Here we cloned the insect neurotoxin BjαIT gene into the pET32 expression vector and expressed the resulting thioredoxin (Trx)-BjαIT fusion protein in Escherichia coli. Soluble Trx-BjαIT was expressed at a high level when induced at 18 °C with 0.1 mM isopropyl β-d-1-thiogalactopyranoside, and it was purified by Ni-nitriloacetic acid affinity chromatography. After cleaving the Trx tag with recombinant enterokinase, the digestion products ... More