猪圆环病毒2型Cap蛋白的纯化与鉴定  

Purification and Identification of Recombinant Porcine Circovirus Type 2 Capsid Protein

作　　者：周景明[1] 贾蕊[1] 刘红亮[1] 祁艳华[1] 聂守一 陈冰[1] 张改平[1] 王爱萍[1] 

机构地区：[1]郑州大学生命科学学院,河南郑州450001

出　　处：《动物医学进展》2017年第9期23-27,共5页Progress In Veterinary Medicine

基　　金：国家重点研发计划项目(2016YFD0500704);郑州市科技创新团队项目(131PCXTD622)

摘　　要：为了获得具有良好免疫原性的猪圆环病毒2型重组Cap蛋白,对pET-28a-ORF2-BL21重组菌种进行IPTG诱导表达,利用镍亲和层析法纯化可溶性重组蛋白,并利用Western blot和ELISA鉴定其免疫反应性。结果表明,表达的重组蛋白分子质量约为26ku,主要以可溶性蛋白的形式存在,纯化后的蛋白纯度可达到90%以上,浓度为1.31mg/mL,产量为30.6mg/L菌液。Western blot和间接ELISA证实,重组蛋白能够与PCV2阳性血清反应。结果表明,所制备的Cap蛋白具有较高的纯度和较好的免疫反应性,有望用于PCV2抗体检测免疫试纸的研制。To obtain the immunogenic PCV2 Cap protein,recombinant bacteria pET-28a-ORF2-BL21 was in-duced with IPTG,the soluble recombinant protein were purified with Ni-affinity chromatography column, then the immunoreactivity of the purified recombinant capsid protein was identified by Western blot and in-direct ELISA. The result showed that soluble recombinant protein with molecular weight of about 26 ku was expressed in recombinant bacteria pET-28a-ORF2-BL21 after induced with IPTG. The purity of puri-fied Cap protein was above 90 % and the concentration was 1. 31 mg/mL. The yield of protein was 30. 6 mg per liter bacteria liquid. Western blot and indirect ELISA showed that the purified protein could react with the PCV2 positive serum. The recombinant Cap protein prepared in this study has high purity and strong immunoreactivity,which has the potential to be used for the development of colloidal gold immunochroma- tographic strip for detection of PCV2 antibody.

关 键 词：猪圆环病毒2型 核衣壳蛋白 蛋白纯化 鉴定 

分 类 号：S852.659.2[农业科学—基础兽医学] Q789[农业科学—兽医学]