西尼罗河病毒NS1的克隆、表达及抗原性分析

Cloning and expression of the NS1 gene encoding the non-structural protein Ⅰ of West Nile Virus and the analysis of its antigenicity

目的克隆和表达西尼罗河病毒(WNV)非结构蛋白1(NS1),并分析其与登革病毒NS1的抗原交叉反应性。方法合成WNV NS1全基因序列,将NS1全基因克隆到pGEX-5X-3原核表达载体中表达GST-NS1融合蛋白,用4型登革病毒免疫血清及登革病毒NS1单克隆抗体并应用Western Blot方法分析WNV-NS1重组蛋白的抗原性。结果重组质粒pGEX-5X-3-WNV-NS1经IPTG诱导,高效表达GST-NS1融合蛋白,经变性纯化和复性获得可溶性WNV-NS1重组蛋白,经Western Blot证实WNV-NS1重组蛋白可与各型登革病毒免疫小鼠血清及部分4型登革病毒NS1交叉单抗识别。结论成功获得可溶性WNV-NS1重组蛋白,该蛋白与登革病毒NS1具有抗原交叉反应性,其结果为建立西尼罗河病毒的血清学诊断和鉴别诊断奠定了基础。

To clone and express the NS1 gene encoding the non-structural protein I of West Nile Virus （WNV） and to analyze the crossing reaction antigens with dengue fever virus, the whole NS1 gene sequence of WNV was synthesized and cloned to prokaryotic expression vector pGEX-SX-3 to express the GST-NS-1 fusion protein. Meanwhile, its antigenicity was analyzed by mouse antisera and monoclonal antibodies against 4 types of NS-1 proteins of dengue fever virus by Western blot assay. Experimental results demonstrated that the recombinant NS-1 protein could be highly expressed in E. coli BL21 and the soluble NS-1 protein could he recognized by the mouse antisera and the part of the crossing monoelonal antibodies against 4 types of NS-1 protein of dengue fever virus. These results would provide foundation for the establishment of the serological diagnosis and the differential diagnosis for WNV infection.