酸弗林蛋白酶酸性氨基酸簇分选蛋白-1的原核表达、纯化及多克隆抗体制备

目的：原核表达和纯化PACS-1，并制备其多克隆抗体。方法：通过RT-PCR扩增出PACS-1的编码基因，测序正确后克隆入原核表达载体pGEX4T-1，转化大肠杆菌BL21（DE3），以IPTG诱导PACS-1与GST融合蛋白的表达并经Glutathione Sepharose 4B纯化；经SDS-PAGE和Western blot鉴定，应用纯化的蛋白免疫家兔制备多克隆抗体，用ELISA测定抗体的效价。结果：表达和纯化了PACS-1，并获得了较高效价的抗血清。结论：获得纯化的PACS-1及其多克隆抗体，为进一步研究PACS-1的功能奠定了基础。

Prokaryotic Expression and Purification of Phosphofurin Acidic Cluster Sorting Protein 1 and Preparation of its Polyclonal Antibody

Objective： To express and purify phosphofurin acidic cluster sorting protein 1 （PACS-1） in E.coli and to prepare rabbit anti-pACS-1 antibody. Methods： PACS-1 gene was amplified by RT-PCR and cloned into prokaryotic expression vector pGEX4T-1. The recombinant vector was transformed into E.coli BI221（DE3）. PACS-1 expression was then induced by IPTG. The expressed GST-PACS-1 fusion protein was purified by Glutathione Sepharose 4B and characterized by SDS-PAGE and Western blot. The purified protein was injected into a rabbit, and the titer of the rabbit＇s anti-serum was measured by ELISA. Results： The PACS-1 was successfully purified and the rabbit＇s anti-serum against PACS-1 with high titer was obtained. Conclusion： The preparation of the purified PACS-1 and its polyclonal antibody can be used for further investigation.