人热休克蛋白gp96基因的克隆及其在大肠杆菌中的表达与纯化

热休克蛋白gp96是热休克蛋白90家族成员，能够引起非特异性和特异性免疫反应。得到大量高纯度的蛋白质是研究开发gp96的关键。然而重组的gp96容易在E．coli中降解，并在一定条件下形成多聚体。实验先将人gp96基因克隆到pET-30a载体上并在E．coli Blstar中表达，再经过亲和层析、阴离子交换、分子筛分别纯化gp96。最终去掉了大部分的降解片段和多聚体，得到一定量的可溶性gp96，为进一步研究其结构和功能打下一定的基础。

Cloning, Expression and Purification of Human Heat Shock Protein GP96 Gene in Escherichia coli

Heat shock protein gp96 is a member of HSP90 families.It can elicits both innate and adaptive immune responses. Generally it is essential to obtain enough amounts of pure gp96 to meet the needs of study and application. But the recombinant gp96 in E.coli is easy to degrade and form aggregates in certain conditions.In the experiment,first cloned human gp96 gene into pET-30a vector and expressed the recombinant in E.coli Blstar. Then purified gp96 by Ni-affinity column,anion exchange column and Gel-filtration in turn.In the end, removes most degraded fragments,aggregates and obtains certain amount of soluble gp96,which make an foundation for further investigations of the protein.