鹅源新城疫病毒F基因的原核表达及间接ELISA诊断方法的建立  

Prokaryotic expression of Newcastle disease virus from goose F gene and development of an indirect ELISA

作　　者：刘新鑫[1] 孟轲音[2] 尹仁福[1] 丁壮[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2008年第12期1383-1386,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30571375,30771606)

摘　　要：参照GenBank公布的鹅源新城疫病毒NA-1株F基因(DQ659677)的核苷酸序列设计1对引物,通过PCR扩增F基因全长,将其克隆到pET-22b(+)载体中,构建了原核表达载体pET-22b(+)-F,转化大肠杆菌BL21(DE3),表达并纯化了重组蛋白,Western-blot证明该重组蛋白具有免疫原性。利用重组蛋白为包被抗原,通过方阵试验确定了包被抗原的最佳包被量为0.21μg/孔,建立了检测鹅源新城疫病毒抗体的间接ELISA诊断方法。According to the F gene sequences of Newcastle disease virus from goose （DQ659677） available in GenBank,a pair of primers was designed for F gene of NDV from goose. Then the amplified product was cloned into prokaryotic expression vector pET-22b（＋） and over-expressed in E. coli BL21 （DE3）. Western-blot analysis showed that the purified recombinant protein had good immunological activity. The optimal concentration of coated antigen F recombinant protein was 0.21ug per well,which was determined by phalanx titrimetry. Then an indirect ELISA was established for detection of the antibody against Newcastle disease virus from goose.

关 键 词：鹅源新城疫病毒 F基因 原核表达 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]