兔出血症病毒衣壳蛋白重组腺病毒的构建、表达及其免疫原性 : = Construction and Expression of Capsid Protein of Rabbit Hemorrhagic Disease Virus Shanghai Strain in the System of Recombinant Adenovirus and Its Immunogenicity in Rabbit
通过RT-PCR方法扩增上海地区兔出血症病毒(RHDV)衣壳蛋白基因(VP60)。将其克隆入腺病毒表达载体系统中,构建成重组腺病毒质粒,用脂质体法转染HEK293细胞,得到重组病毒pAd-VP60,经RT-PCR、IFA、HA、SDS-PAGE和Western-blotting鉴定,结果表明重组VP60蛋白在HEK293细胞中获得表达。表达产物与铝胶佐剂混合后注射3月龄非免疫健康家兔,结果表明,免疫后21 d兔体内产生抗RHDV抗体,其HI效价可达2~6～2~7,能抵御HA≥2~(10)致死剂量RHDV强毒的攻击,保护率为100%。.
The capsid protein VP60 gene of rabbit hemorrhagic disease virus was amplified by revere transcript polymerase chain reaction(RT-PCR) technique.The fragment was cloned into the system of adenovirus expression and trans- fected into HEK293 cells,and a recombinant adenovirus named pAd-VP60 was obtained.The expressed protein was identified by RT-PCR,IFA,HA,SDS-PAGE and Western-blotting.The recombinant protein of VP60 was expressed in HEK293 cells.The expressed protein was mixed with aluminum rubber adjuvant and applied to immunize three-month-old rabbits without immunized RHD vaccine.The results indicated that the anti-RHDV antibodies were induced and developed after 21 d of immunization, with the HI titers of serum ranging from 2~6 to 2~7. When challenged with HA≥2~(10) units of infectious RHDV,the control rabbits died while the immunized animals all survived.The results suggested that the recombinant proteins expressed by recombinant adenovirus could be promising candidates for RHDV live vaccines..
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2009-12-31 2009 |
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| Erschienen: |
2009-12-31 |
| Enthalten in: |
Zur Gesamtaufnahme - year:2009 |
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| Enthalten in: |
Jiang su nong ye xue bao - (2009), 06 vom: 31. Dez., Seite 1320-1324 Original Letters: Enthalten in 江苏农业学报 (DE-600)2993832-6 (DE-600)2993832-6 江苏省南京市 |
| Reihe: |
China Academic Journals (CAJ), D, 农业科技 = Agriculture China Academic Journals (CAJ), A, 理工A(数学物理力学天地生) = Mathematics/ Physics/ Mechanics/ Astronomy |
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| Sprache: |
Chinesisch |
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| Weiterer Titel: |
Construction and Expression of Capsid Protein of Rabbit Hemorrhagic Disease Virus Shanghai Strain in the System of Recombinant Adenovirus and Its Immunogenicity in Rabbit |
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| Beteiligte Personen: |
胡波 [VerfasserIn] |
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| Links: |
oversea.cnki.net [lizenzpflichtig] |
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| Anmerkungen: |
Author info:HU Bo~(1,2),WANG Fang~1,QIN Ai-jian~2,FAN Zhi-yu~1,XU Wei-zhong~1,ZHANG Ze-bin~1,XUE Jia-bin~1, HE Kong-wang~1 (1.Institute of Veterinary Medicine Jiangsu Academy of Agricultural Sciences,Key Laboratory of Animal Diseases Diagnosis and Immunology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;2.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China) |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
CAJ322842255 |
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| 245 | 1 | 0 | |a 兔出血症病毒衣壳蛋白重组腺病毒的构建、表达及其免疫原性 |b = Construction and Expression of Capsid Protein of Rabbit Hemorrhagic Disease Virus Shanghai Strain in the System of Recombinant Adenovirus and Its Immunogenicity in Rabbit |
| 246 | 3 | 1 | |a Construction and Expression of Capsid Protein of Rabbit Hemorrhagic Disease Virus Shanghai Strain in the System of Recombinant Adenovirus and Its Immunogenicity in Rabbit |
| 264 | 1 | |c 2009-12-31 | |
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| 490 | 0 | |a China Academic Journals (CAJ) |a A |a 理工A(数学物理力学天地生) = Mathematics/ Physics/ Mechanics/ Astronomy | |
| 500 | |a Author info:HU Bo~(1,2),WANG Fang~1,QIN Ai-jian~2,FAN Zhi-yu~1,XU Wei-zhong~1,ZHANG Ze-bin~1,XUE Jia-bin~1, HE Kong-wang~1 (1.Institute of Veterinary Medicine Jiangsu Academy of Agricultural Sciences,Key Laboratory of Animal Diseases Diagnosis and Immunology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;2.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China) | ||
| 520 | |a 通过RT-PCR方法扩增上海地区兔出血症病毒(RHDV)衣壳蛋白基因(VP60)。将其克隆入腺病毒表达载体系统中,构建成重组腺病毒质粒,用脂质体法转染HEK293细胞,得到重组病毒pAd-VP60,经RT-PCR、IFA、HA、SDS-PAGE和Western-blotting鉴定,结果表明重组VP60蛋白在HEK293细胞中获得表达。表达产物与铝胶佐剂混合后注射3月龄非免疫健康家兔,结果表明,免疫后21 d兔体内产生抗RHDV抗体,其HI效价可达2~6～2~7,能抵御HA≥2~(10)致死剂量RHDV强毒的攻击,保护率为100%。 | ||
| 520 | |a The capsid protein VP60 gene of rabbit hemorrhagic disease virus was amplified by revere transcript polymerase chain reaction(RT-PCR) technique.The fragment was cloned into the system of adenovirus expression and trans- fected into HEK293 cells,and a recombinant adenovirus named pAd-VP60 was obtained.The expressed protein was identified by RT-PCR,IFA,HA,SDS-PAGE and Western-blotting.The recombinant protein of VP60 was expressed in HEK293 cells.The expressed protein was mixed with aluminum rubber adjuvant and applied to immunize three-month-old rabbits without immunized RHD vaccine.The results indicated that the anti-RHDV antibodies were induced and developed after 21 d of immunization, with the HI titers of serum ranging from 2~6 to 2~7. When challenged with HA≥2~(10) units of infectious RHDV,the control rabbits died while the immunized animals all survived.The results suggested that the recombinant proteins expressed by recombinant adenovirus could be promising candidates for RHDV live vaccines. | ||
| 610 | 2 | 4 | |a 江苏省农业科学院兽医研究所,农业部动物疫病诊断与免疫重点开放实验室,国家兽用生物制品工程技术研究中心 |
| 610 | 2 | 4 | |a 扬州大学兽医学院 |
| 650 | 4 | |a 动物医学（兽医学） | |
| 650 | 4 | |a 畜牧、动物医学、狩猎、蚕、蜂 | |
| 650 | 4 | |a 农业科学 | |
| 650 | 4 | |a Biology | |
| 650 | 4 | |a Animal Husbandry and Veterinary | |
| 650 | 4 | |a 农业科技 | |
| 650 | 4 | |a Agriculture | |
| 650 | 4 | |a 理工A(数学物理力学天地生) | |
| 650 | 4 | |a Mathematics/ Physics/ Mechanics/ Astronomy | |
| 700 | 0 | |a 王芳 |4 oth | |
| 700 | 0 | |a 秦爱建 |4 oth | |
| 700 | 0 | |a 范志宇 |4 oth | |
| 700 | 0 | |a 徐为中 |4 oth | |
| 700 | 0 | |a 张则斌 |4 oth | |
| 700 | 0 | |a 薛家宾 |4 oth | |
| 700 | 0 | |a 何孔旺 |4 oth | |
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