使用TEV-LIC克隆与表达系统对蛋白激发子PebC1的纯化

摘要

PebC1是来自于灰葡萄孢菌（Botrytis cinerea）的蛋白激发子,具有提高植物综合抗逆性的功能,可以开发为生物农药。由于该蛋白自身性质原因,如结合杂蛋白、形成二聚体等,难于得到纯蛋白,不利于进行进一步的研究。将该蛋白的编码基因克隆到TEV-LIC表达载体系统,并对该蛋白进行了表达,使用His标签和MBP标签进行亲和层析,然后通过TEV蛋白酶切去标签,用离子交换层析和分子筛层析得到高纯度的蛋白。本研究为PebC1的后续研究打下了基础,也为研究一些难于纯化的蛋白提供了新思路。%PebC1 is a protein elicitor from Botrytis cinerea.This protein can promote the overall stress tolerance of plant and can be developed as biological pesticide.But it was difficult to get purified protein for its own characters like binding peptides,forming homodimer etc,which would help forfurther study of PebC1.In this research,coding gene was cloned into a TEV-LIC system for expression.The fusion protein was purified by His and MBP tags affinity chromatography,then the tags was cut off by TEV protease.Finally,the high purity PebC1 was separated by ion-exchange and size-exclusive chromatography.This research laid foundation for the further research on PebC1 and shed new light on the research of proteins hard to be purified.