| 鸭坦布苏病毒E基因在昆虫细胞中的分泌表达及免疫原性与保护性研究 |
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| 引用本文： | 王善辉,谢金文,吕颜枝,李云龙.鸭坦布苏病毒E基因在昆虫细胞中的分泌表达及免疫原性与保护性研究[J].中国畜牧兽医,2015,42(10):2580-2586. |
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| 作者姓名： | 王善辉 谢金文 吕颜枝 李云龙 |
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| 作者单位： | 1. 江苏省徐州生物工程职业技术学院动物工程系, 徐州 221006;2. 山东省滨州畜牧兽医研究院重点实验室, 滨州 256600 |
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| 基金项目： | 徐州市科技项目(XM13B120);徐州生物工程职业技术学院科研课题(2013B03) |
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| 摘 要： | 鸭坦布苏病毒(DTMUV)为新出现的病毒,主要引起鸭产蛋量急剧下降。本研究通过杆状病毒表达系统,利用蜂素信号肽(honeybee melittin signal peptide,Mels)构建分泌表达DTMUV JSXZ株E蛋白的重组杆状病毒,将其感染Sf9昆虫细胞,从而分泌表达DTMUV E蛋白,并对该表达E蛋白的免疫原性和保护效果进行了研究。经Western blotting、IFA试验证明E蛋白可以在Sf9昆虫细胞培养上清中分泌表达;通过免疫雏鸭试验、间接ELISA结果证明E蛋白可以诱导产生高水平的IgG血清抗体;MTT试验结果显示E蛋白能刺激T 淋巴细胞增殖;攻毒保护试验结果显示0.3 mL组免疫保护率为80%,0.6 mL组和0.8 mL组保护率均为100%,而PBS对照组雏鸭均表现为典型的DTMUV感染症状,其中3只死亡。上述研究结果为DTMUV亚单位疫苗的研究奠定基础。
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| 关 键 词： | 鸭坦布苏病毒(DTMUV) E蛋白 杆状病毒 分泌表达 免疫原性 保护效果 |
| 收稿时间： | 2015-02-05 |
Secretory Expression of DTMUV E Gene in Insect Cells and Study on its Immunogenicity and Protective Efficacy |
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| WANG Shan-hui,XIE Jin-wen,LV Yan-zhi,LI Yun-long.Secretory Expression of DTMUV E Gene in Insect Cells and Study on its Immunogenicity and Protective Efficacy[J].China Animal Husbandry & Veterinary Medicine,2015,42(10):2580-2586. |
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| Authors: | WANG Shan-hui XIE Jin-wen LV Yan-zhi LI Yun-long |
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| Institution: | 1. Animal Engineering Department, Jiangsu Xuzhou Vocational College of Bioengineering, Xuzhou 221006, China;2. Key Laboratory of Shandong Binzhou Animal Science & Veterinary Medicine Institute, Binzhou 256600, China |
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| Abstract: | Duck Tembusu virus (DTMUV) was newly emerged which caused severe egg-drop syndrome in duck.Here,we constructed the recombinant baculovirus by honeybee melittin signal peptide (Mels) which expressed E protein of DTMUV JSXZ strain,and its immunogenicity and protective efficacy were investigated in duck infection model.Western blotting and indirect immunofluorescence assay (IFA) showed E protein was effectively secretory expressed in supernatant of Sf9 insect cells.Indirect ELISA result showed E protein could induce high levels of anti-E IgG in the sera.MTT result showed that E protein could stimulate the proliferation of T lymphocytes.Moreover,the protection rates were 80% in 0.3 mL group and 100% in both 0.6 mL and 0.8 mL groups,respectively,whereas,all ducks in PBS control group showed the typical clinical signs with 3 died post challenge.These results laid a foundation for the study of DTMUV subunit vaccine. |
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| Keywords: | duck Tembusu virus (DTMUV) E protein baculovirs secretory expression immunogenicity protective efficacy |
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