摘要
利用生物软件对新城疫病毒F48E9株的血凝素神经氨酸酶(HN)蛋白进行抗原特性分析,选定172～570位氨基酸区域作为多肽表位候选区域。以pUC18-F-HN为模板,设计引物通过PCR扩增,获得HN抗原结构域基因片段,SaⅠl、NoⅠt双酶切定向克隆到原核表达载体pET28a,获得重组质粒分别命名为pET28a-HNa。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,HN抗原结构域基因片段获得了融合表达,Western-blotting分析证实表达产物HN与NDV阳性血清具有免疫反应性。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蛋白相互作用奠定了基础。
The antigenicity of hemagglutinin-neuraminidase（HN） protein in Newcastle Disease Virus（NDV） strain F48E9 was analyzed by using biological software,the section of 172～570 AA was selected as candidates for peptide epitopes.The gene fragments HN coding for the antigenic structural domains of NDV HN were amplified from the recombinant plasmid containing full length of NDV HN gene by PCR with specific primers.The genes were inserted between SalⅠand NotⅠsites of pET28a after cleavage by corresponding enzymes.The recombinant plasmids were named as pET28a-HNa.It was found that the recombinant plasmids could be expressed in E.coli BL21 and the expressed fragments HN could be recognized by NDV positive serum from SPF chickens through Western blotting analysis.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.
出处
《江西农业学报》
CAS
2010年第7期91-94,共4页
Acta Agriculturae Jiangxi
基金
江西省科技支撑计划(2009BNA06600
2008BC34400)
江西省科学院国家级预研项目(2008-1-01)
关键词
新城疫病毒
血凝素-神经氨酸酶
结构域
原核表达
Newcastle disease virus
Hemagglutinin-neuraminidase
Structural domains
Prokaryotic expression
