Ⅰ型鸭肝炎病毒VP3基因的原核表达及其抗原性

摘要

参考GenBank 中的Ⅰ型鸭肝炎病毒(DHV Ⅰ)全基因序列设计引物,采用RT-PCR方法扩增了DHV Ⅰ VP3 基因,将VP3 基因与pET-28a(+)表达载体连接后,转化宿主BL21(DE3),用0.1 mmol·L-1的IPTG诱导表达,收集菌液进行SDS-PAGE电泳,Western-blotting分析蛋白免疫原性.电泳结果表明,VP3在大肠杆菌中表达量较高,表达产物的分子量约为27 kD.免疫印迹试验表明 Ⅰ型鸭肝炎病毒VP3蛋白在大肠杆菌中表达产物具有免疫原性.%In this study, a pair of special primers, according to the complete genome of Duck hepatitis virus Ⅰ ( DHV- Ⅰ ) , was designed to amplify VP3 gene of DHV- Ⅰ . VP3 was subcloned into pET-28a( + ), then the recombinant plasmids were transformed into Escherichia coli BL21 ( DE3 ) and induced by 0. 1 mmol· L-1 IPTG. The bacteria containing pET-28a( + )VP3 were collected and examined by SDS-PAGE and Western-blotting. The results showed that VP3 protein was well expressed in E. coli. The molecular weight of the recombinant protein was 27 kD. The protein could be recognized by the specific antibody against DHV- Ⅰ .