H5亚型禽流感病毒样颗粒的构建及生物学活性鉴定  

Construction and identification of H5 subtype avian influenza virus-like particles

作　　者：张超林[1,2] 刘春国[2] 王寿山[2] 石薇琳[2] 李建辉[2] 王伟[1,2] 刘彦云[2] 刘明[1,2] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2012年第7期514-517,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：现代农业技术体系(CARS-43-10);黑龙江省自然科学基金(ZJN0702-02)

摘　　要：为构建H5亚型禽流感病毒(AIV)病毒样颗粒(VLPs),并鉴定其生物学活性,本研究通过杆状病毒/昆虫细胞表达系统,分别单独表达H5亚型AIV血凝素(HA)及共表达HA和基质蛋白(M1),并采用免疫组织化学法、SDS-PAGE、western blot和血凝试验等对重组蛋白进行鉴定。实验结果表明:透射电子显微镜观察结果显示,共表达HA和M1的重组蛋白可以形成VLPs,而单独表达HA未观察到VLPs;表达的重组蛋白HA和HA-M1均能够凝集鸡红细胞,血凝效价分别为1∶27和1∶28。结果显示该方法获得的AIV结构蛋白具有良好的生物学活性和抗原活性,从而为禽流感VLPs疫苗的研究提供依据。To prepare the virus-like particles （VLPs） of H5 subtype avian influenza virus （AIV）, the recombinant baculovirus expressing the hemagglutinin （HA） or both HA and matrix 1 （M1） were constructed and expressed in Sf9 cells, and the bioactivities of VLPs were identified by immunohistochemistry and hemagglutination test. The SDS-PAGE and western blot analysis indicated that the HA and HA-M1 were efficiently expressed and the VLPs were observed under transmission electron microscopy in Sf9 cells, which infected only with the recombinant baculovirus co-expressing HA and M1 of H5 subtype A1V. The immunohistochemistry and hemagglutination test demonstrated that both of the HA and HA-M1 expressed in the cells and the HA titers were 27 and 28, respectively. The VLPs produced in Sf9 cells facilitate for the further study and development of the novel avian influenza vaccines.

关 键 词：流感病毒 血凝素基因 基质蛋白基因 杆状病毒表达系统 

分 类 号：S852.65[农业科学—基础兽医学]