麻疹病毒血凝素和融合蛋白基因的克隆与表达及其重组蛋白的免疫学评价  

Cloning and expression of hemagglutinin and fusion protein of measles virus

作　　者：唐凌云[1] 张鹏飞[1] 章以浩[1] 

机构地区：[1]卫生部北京生物制品研究所

出　　处：《中华微生物学和免疫学杂志》1997年第4期261-266,共6页Chinese Journal of Microbiology and Immunology

摘　　要：采用逆转录－多聚酶链反应（ＲＴ－ＰＣＲ）的方法从麻疹病毒Ｅｄｍｏｎｓｔｏｎ株基因组中扩增出血凝素Ｈ基因和融合蛋白Ｆ基因，并利用转移质粒将这两个基因分别重组到杆状病毒多角体蛋白启动子（ＰＨ）控制之下，获得重组病毒ｖＢＭＶＨ和ｖＢＭＶＦ。重组杆状病毒感染Ｓｆ９昆虫细胞，表达的重组蛋白分别具有血凝、血溶活性。红细胞吸附抑制试验、免疫印迹、酶联免疫试验结果显示重组蛋白在生物学、生化学特性与天然蛋白类似，并能被天然麻疹免疫血清（人、鼠、兔）所识别。动物实验表明，重组蛋白具有诱生中和抗体的能力。重组血凝素免疫血清还具有血凝抑制抗体活性。这些结果说明杆状病毒－昆虫细胞体系表达的麻疹病毒血凝素和融合蛋白具有较好的生物学活性及免疫原性和抗原性。The genes coding for hemagglutinin(H) and fusion protein(F) of measles virus were obtained from viral RNA extracted from supernatant of viral culture by RT PCR,and were cloned into baculovirus transfer vectors,respectively.Sf9 cells were co transfected with recombinant vectors and wild type baculovirus DNA,and two recombinant baculoviruses containing the genes of H and F were isolated through several rounds of plaque purification.Recombinant protein H and recombinant protein F were expressed in Sf9 insect cells post infected with recombinant baculoviruses evidenced by recognition of recombinant proteins with anti sera in both Western blot and ELISA.Hemagglutination and hemolysis of recombinant proteins have been shown by recombinant baculoviruses infected cell lysates.The results of immunization assay with recombinant baculoviruses infected cell lysates indicated that the recombinant proteins expressed in insect cells had the ability of elicitation of specific antibodies against native measles virus in mice suggesting that recombinant H and recombinant F expressed in insect cells had as same biological properties as their native counterparts.

关 键 词：麻疹病毒 融合蛋白 血凝素 杆状病毒 基因表达 

分 类 号：R511.103[医药卫生—内科学]