【摘要】 鸭坦布苏病毒(DTMUV)为新出现的病毒,主要引起鸭的产蛋量急剧下降。为评价真核表达DTMUV的E蛋白的免疫原性,本研究利用杆状病毒表面展示技术,构建表面展示DTMUV JSXZ株E蛋白的重组杆状病毒(r BV-DTMUV-E),将其感染昆虫细胞,从而将E蛋白展示在杆状病毒的表面,并对表达的重组E蛋白(r E)的免疫原性和保护效果进行研究。经western blot和间接免疫荧光试验证明r E能够在Sf9细胞中有效表达;并且呈可溶性表达。重组蛋白经镍柱纯化,并采用氢氧化铝乳化(0.1μg/m L),分别以0.2 m L、0.4 m L和0.6 m L的剂量免疫雏鸭,通过ELISA抗体检测结果证明,r E可以诱导产生高水平的Ig G血清抗体;MTT试验结果表明,r E能够刺激T淋巴细胞增殖;攻毒保护试验结果显示,其0.2 m L组免疫保护率为80%,0.4 m L和0.6 m L组保护率均为100%,而对照组雏鸭均表现为典型的DTMUV感染症状,其中4只死亡。以上研究结果为DTMUV亚单位疫苗的研究奠定了基础。更多还原

【Abstract】 Duck Tembusu virus(DTMUV) was newly emerged virus which causes severe egg-drop syndrome in duck. In the study, a recombinant baculovirus(r BV-DTMUV-E) contained E gene of DTMUV JSXZ stain was constructed by baculovirus surface-display technology, and the recombinant E protein(r E) was expressed and evaluated for the immunoprotection in ducklings.Western blot and indirect immunofluorescence assay showed that the r E protein was effectively expressed in r BV-DTMUV-E infected Sf9 cells in a soluble form. Subsequently, the r E was purified with nickel-chelate resin and emulsified with with aluminium hydroxide to immunize ducklings at a dosage of 0.1, ELISA detection results showed the high level of antibody was induced in the sera from the r E protein immunized ducklings. MTT results showed that r E protein was able to stimulate the proliferation of T lymphocytes. Moreover, the protection rates of the r E protein were 75% in 0.2 m L group and 100% in both 0.4 m L and 0.6 m L groups, respectively, whereas, all duckings in control group shown the typical clinical signs with 4 died postchallenge.These results provided a basis for further study on DTMUV subunit vaccine.更多还原