摘要
[ Objective ] This paper aimed to obtain active alpha-bungarotoxin （α-BGT） protein and investigate the isolation and purification of recombinant α-BGT protein. [ Method] The expressed soluble fusion protein was purified by using GSTrap FF affinity columns. Purified fusion protein bound to GSTrap FF affinity col- umns was directly cleaved by thrombin to obtain the solution containing recombinant α-BGT. Using natural α-BGT as control, the antigenicity and biological activity of the purified fusion protein and recombinant α-BGT were detected by SDS-PAGE, Western Blot and toxicity test in vivo. [ Result] Recombinant ot-BGT was iso- lated; the purified fusion protein and recombinant α-BGT were similar to the natural α-BGT in antigenicity; the toxicity of purified fusion protein was relatively wea- ker; recombinant α-BGT was similar to the natural ot-BGT in toxicity. [Condusion] This study laid the foundation for further large-scale production of recombi- nant α-BGT.
[ Objective ] This paper aimed to obtain active alpha-bungarotoxin （α-BGT） protein and investigate the isolation and purification of recombinant α-BGT protein. [ Method] The expressed soluble fusion protein was purified by using GSTrap FF affinity columns. Purified fusion protein bound to GSTrap FF affinity col- umns was directly cleaved by thrombin to obtain the solution containing recombinant α-BGT. Using natural α-BGT as control, the antigenicity and biological activity of the purified fusion protein and recombinant α-BGT were detected by SDS-PAGE, Western Blot and toxicity test in vivo. [ Result] Recombinant ot-BGT was iso- lated; the purified fusion protein and recombinant α-BGT were similar to the natural α-BGT in antigenicity; the toxicity of purified fusion protein was relatively wea- ker; recombinant α-BGT was similar to the natural ot-BGT in toxicity. [Condusion] This study laid the foundation for further large-scale production of recombi- nant α-BGT.
