摘要
目的克隆猪囊尾蚴胞质含铜/锌的超氧化物歧化酶基因(Cu/ZnSOD),并比较其与其它寄生蠕虫相应基因结构的相似性。方法通过获取其它生物Cu/ZnSOD基因的保守区域,设计保守引物,用于扩增该寄生虫的Cu/ZnSOD基因。结果Cu/ZnSOD基因编码一15.6kDa的蛋白,该蛋白的推导序列中含有这类酶的活性和二级结构所需要的所有保守的氨基酸残基,并且与其它寄生虫的相应序列有高达70.6%的相似度。抑制剂研究试验表明所克隆的SOD属于Cu/ZnSOD,该蛋白在大肠杆菌中得以成功表达,表达产物具有SOD活性。免疫印迹试验发现猪囊尾蚴Cu/ZnSOD对人不仅具有抗原性,而且与其它几种蠕虫的Cu/ZnSOD有广泛的交叉免疫反应。结论克隆并表达出猪囊尾蚴Cu/ZnSOD,它与其它寄生吸虫和绦虫的Cu/ZnSOD在氨基酸水平具有很高的相似度,因此是理想的广谱抗寄生虫药物和疫苗的靶蛋白,值得进一步研究。
A copper/zinc superoxide dismutase （Cu/Zn SOD） gene was cloned and its gene product was functionally characterized. The conserved primers were designed via the Cu/Zn SOD gene of other species and used to amplify the Cu/Zn SOD gene of this parasite. A 15.6 kDa protein encoded by the Cu/Zn SOD gene was obtained. The deduced amino acid sequence of the gene product contained highly conserved residues required for activity and the second sturcture formation of Cu/ Zn SOD, and it showed up to 70.6 % of identity in the corresponding sequences with their counterparts in other animals. Experiments with inhibitors indicated that the cloned SOD belonged to the Cu/Zn SOD, which was functionally expressed in E. coli and exhibited SOD activity. Result in immunoblotting showed that the Cu/Zn SOD of T. soliurn was antigenic to humans and demonstrated extensive cross-reaction with sera of patients infected with other trematodes or cestodes, indicating that this protein may be the ideal target for the broad-spectrum anti-parasitic drugs and vaccines.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2006年第12期1124-1128,共5页
Chinese Journal of Zoonoses
基金
韩国健康与社会福祉部博士后基金资助(NIH-348-6111-211)
