重组敬钊毒素在毕赤酵母中的表达与纯化研究

The Expression and Purification of Jingzhao Toxin in Pichia pastoris

[目的]研究重组敬钊毒素在毕赤酵母中的表达与纯化。[方法]通过设计、构建Jztx-III/pPIC9k表达载体,经电转化、甲醇诱导后使敬钊毒素在毕赤酵母GS115中进行表达,并利用C-端6×His标签进行分离纯化。[结果]Jztx-III可以在毕赤酵母GS115中高效表达,经亲和层析纯化后其产量可达到95 mg/L。[结论]该试验结果解决了敬钊毒素资源有限产量较低的问题。

[Objective] To study expression and purification of Jingzhao toxin in Pichia pastoris.[Method] Jztx-Ⅲ GS115/pPIC9k plasmid was constructed to express Jingzhao toxin-Ⅲ in Pichia pastoris.A 6 × His tag in C-terminus was employed to purify the recombinant Jztx-Ⅲ.[Result] The results showed that Jztx-Ⅲ can be expressed in Pichia pastoris GS115 effectively.The production of Jztx-Ⅲ can reach to 95 mg/L after purified by affinity chromatography.[Conclusion] The experiment results solve the problem of lower yield of Jingzhao toxin limited resource.