新城疫病毒HN蛋白抗原表位分析及结构域基因原核表达  

Prokaryotic expression of structural domains of the newcastle disease virus hemagglutinin-neuraminidase

作　　者：陈亚波[1] 徐程[1] 张桂芝[1] 方维焕[1] 

机构地区：[1]浙江大学动物预防医学研究所浙江省动物预防医学重点实验室,浙江杭州310029

出　　处：《中国预防兽医学报》2007年第1期32-35,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：浙江省自然科学基金重大项目(ZA0105)

摘　　要：以本实验室构建的含新城疫病毒血凝素-神经氨酸酶基因重组质粒为模板,设计引物通过PCR和重叠-延伸PCR扩增,获得HNa、HNb和HNa-L-b三种抗原结构域基因片段,BamHⅠ/HindⅢ双酶切定向克隆到原核表达载体pET32c,获得重组质粒分别命名为pET32c-HNa、pET32c-HNb和pET32c-HNa-L-b。重组质粒转化大肠杆菌感受态细胞BL21,筛选出阳性克隆,诱导表达并取产物进行分析。结果表明,HNa、HNb、HNa-L-b结构域基因片段均获得了融合表达。Western-blotting分析证实表达产物HNa和HNb与NDV阳性血清具有免疫反应性。本试验结果为进一步研究HN蛋白抗原结构域的免疫原性以及HN蛋白与F蛋白在细胞融合中的相互作用奠定了基础。The gene fragments HNa, HNb and HNa-L-b coding for the antigenic structural domains of the newcastle disease virus （NDV） hemagglutinin-neuraminidase were amplified fi＇om the recombinant plasmid containing full length NDV FIN gene by regular PCR or overlap extension PCR with specific primers. The genes were inserted between BamH Ⅰ and Hind Ⅲ sites of pET32c after cleavage by corresponding enzymes. The recombinant plasmids were named pET32c-HNa, pET32c-HNb and pET32c-HNa-L-b. It was found that all recombinant plasmids can be expressed in E. coli BL21 and the expressed fi＇agments HNa and HNb can be recognized by NDV positive serum fi＇om SPF chickens using western-blotting. This study established a foundation for the further examination of the antigenicity of these antigenic structural domains as well as their co-reaction with F protein in cell fusion.

关 键 词：新城疫病毒 血凝素-神经氨酸酶 结构域 抗原表位 原核表达 

分 类 号：S852.659.5[农业科学—基础兽医学] Q78[农业科学—兽医学]