摘要
根据GenBank上发表的犬细小病毒(CPV)VP2蛋白基因序列,设计并合成1对引物,通过PCR扩增VP2基因全长。将其克隆到pET-30a载体中,构建了原核表达载体pET-VP2,转化大肠杆菌Rosetta,表达并纯化了重组蛋白,Western-blotting证明该重组蛋白具有免疫原性。利用重组蛋白为抗原,通过方阵试验确定了包被抗原的最佳包被量为15.8 ng/孔,血清的最佳稀释倍数为1∶100,建立了检测CPV血清抗体的间接ELISA方法。
According to the published canine parvovirus（CPV） VP2 gene in GenBank,a pair of primers was designed and synthesized to amplify the VP2 gene. The amplified product was cloned into expression vector pET-30a and over-expressed in E. coli Rosetta. Western-blot analysis showed the purified recombinant protein had good immunological activity. The optimal concentration of coated antigen VP2 recombinant protein was 15.8 ng per well and the optimal dilution of serum was 1 ： 100, which was determined by phalanx titrimetry. Then an indirect ELISA was established to detect the antibody against CPV.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第3期218-222,共5页
Chinese Veterinary Science
基金
中央级科研院所科技基础性工作专项资金项目(2001DEB20055)
