新城疫病毒HN基因主要抗原位点区段的原核表达

Prokaryotic Expression of Antigen Sites of the Newcastle Disease Virus Hemagglutinin-Neuraminidase

利用生物软件对新城疫病毒GD/GM/03/Ch的血凝素神经氨酸酶（HN）蛋白进行抗原特性分析,选定169-267位氨基酸、318-405位氨基酸和448-571位氨基酸3个区域作为多肽表位候选区域.用含新城疫病毒HN基因的重组质粒为模板,设计引物通过PCR扩增获得HNa、HNb和HNc 3个抗原结构域基因片段,分别经双酶切定向克隆到原核表达载体pBT上进行表达,并构建了pBT-HNa-b-c串联重组质粒进行表达,经过SDS-PAGE和W est-ern-b lot分析,验证了表达产物具有反应原性.

Antigenicity analysis of Newcastle disease virus （NDV） strain GD/GM/03/Ch hemagglutininneuraminidase（HN） protein was studied by using biological software, the selection of 169 -267AA,318 -405AA and 448 -571AA as candidates for peptide epitopes was determined. The gene fragments HNa, HNb and HNc coding for the antigenic structural domains of the NDV HN amplified from the recombinant plasmid containing full length NDV HN gene by regular PCR with specific primers. The genes were respectively inserted into pBT vector after cleavage by corresponding enzymes. The recombinant plasmids pBT-HNa-b-c were also constructed to carry out the expression. It was found that all the expression products were recognized by NDV positive serum in Western-blot test.