中文摘要：

目的：表达GST-ataxin-3-N融合蛋白并制备GST—ataxin-3特异性抗体，为深入研究其功能及其在SCA3发病机制中的作用提供重要的技术和材料保障。方法：将人ataxin-3氨基端基因克隆入原核表达载体pGEX-4T-2，在大肠杆菌（E．coli）BL21中表达，用Glutathione sepharose4B凝胶亲和柱纯化目的蛋白。利用纯化的GST—ataxin-3-N蛋白制备多克隆抗体。结果：成功构建了原核表达载体，得到高表达量的融合蛋白，经亲和层析柱纯化获得较高纯度的GST—ataxin-3-N融合蛋白。以融合蛋白免疫新西兰免得到Ataxin-3-N多克隆抗体，Western Blotting及免疫荧光均证实该抗体能够识别Ataxin-3-myc蛋白，具有较高特异性。结论：利用原核表达人GST-ataxin-3-N融合蛋白制备的Ataxin-3多克隆抗体具有较好的特异性，可用于该蛋白的相关研究。

英文摘要：

Objective： To obtain ataxin-3 fusion protein and prepare anti-ataxin-3 polyclonal antibody. Methods： The DNA fragments encoding N-terminal 177 amino acids of ataxin-3 was amplified by PCR and cloned into a prokaryotic expression vector, pGEX-4T-2, and transformed into E.coli BL21. The expressed proteins were purified from total proteins with Glutathione sepharose 4B agarose column. The New Zealand rabbits were immunized with the purified fusion proteins to prepare polyclonal antiserum. Results： Prokaryotic expression vectors of ataxin-3 fragements were successfully constructed, the target proteins were expressed efficiently after IPTG induction. Western blotting and Immunoflurescence analysis suggest that the polyclonal antibody raised in the rabbit could recognize Ataxin-3-myc and GST protein in human cells. Conclusion： The antiserum has the advantage of good specificity and sensitive affinities.