产气荚膜梭菌ε毒素细胞作用受体的分离与鉴定

产气荚膜梭菌ε毒素（Epsilon toxin，ETX）是B、D型产气荚膜梭菌产生一种的主要致病因子，可导致羊、牛及羔羊和犊牛等动物的致死性肠道疾病，给我国乃至世界畜牧养殖业生产造成严重损失，因此，加强产气荚膜梭菌ETX致病机制的研究尤为必要。虽然ETX的晶体结构得到了详尽的解析，然而ETX在细胞表面的受体一直没有得到确定，严重阻碍了关于ETX入侵细胞机制和治疗方法的研究。本课题以ETX和MDCK细胞为研究对象，通过免疫共沉淀方法分离ETX在MDCK细胞上的受体，然后采用串联质谱技术对分离的蛋白进行鉴定，再通过基因克隆、基因敲除、表面等离子体共振等一系列技术对ETX和受体的相互作用进行充分的论证和研究。本课题的顺利实施有望鉴定出ETX在MDCK细胞表面的受体，这对阐明ETX致病机制和探索新的治疗靶点具有重要意义。

Epsilon toxin (ETX) produced by strains of Clostridium perfringens types B and D is an aetiological agent of animal (goats, sheep, cattle, lambs and calves) enteric illness that can be rapidly fatal and economically devastating at home and abroad. Therefore, it is essential to enhance the study of pathogenic mechanism of ETX. The crystal structure of ETX was resolved, however the receptor of ETX was still not known, which blocks the progress of the investigation of mechanism of ETX binding a cell and therapeutics. Here in this project, the receptor of ETX will be isolated from MDCK cells using co-immunoprecipitation, and be identified by HPLC-MS/MS technology. Gene manipulation technique, gene knock-out, and surface plasmon resonance was used in this study to clarify the interactions between ETX and it’s receptor. The implementation of the project is expected to identify the ETX receptor, which is of important significance to clarify the action mechanism of ETX and explore a new therapeutic target.