| 逆转录病毒载体介导的猪瘟病毒E2基因在真核细胞中表达及动物免疫试验 |
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| 作者姓名： | 许信刚 胡建和 张彦明 |
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| 作者单位： | 西北农林科技大学动物科技学院 陕西杨凌712100
(许信刚)，河南科技学院动物科学系 河南新乡453003
(胡建和)，西北农林科技大学动物科技学院 陕西杨凌712100(张彦明) |
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| 基金项目： | 国家自然科学基金项目(30270988)。 |
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| 摘 要： | 利用DNA重组技术将猪瘟病毒(CSFV)石门株囊膜蛋白E2基因插入逆转录病毒载体pBABE-puro中构建成重组逆转录病毒载体pBABE-puro-E2,该重组逆转录病毒表达载体与pVSVg质粒经磷酸钙共转染法将其转入293GP细胞中包装逆转录病毒假病毒。用包装的假病毒感染SP2/0细胞,经嘌呤霉素筛选阳性细胞后进行流式细胞技术(FACS)分析,结果表明CSFVE2基因在SP2/0细胞膜上成功表达。将表达E2蛋白的SP2/0细胞腹腔免疫BALB/c小鼠,用流式细胞仪检测证明,成功诱导小鼠产生了抗E2蛋白的抗体。为下一步利用细胞免疫小鼠研制抗猪瘟病毒单克隆抗体打下基础。
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| 关 键 词： | 猪瘟病毒 E2囊膜蛋白 逆转录病毒载体 FACS |
Retroviral Vector Mediated CSFV E2 Envelope Protein Gene Expression Eukaryotic Cells and Animal Immune Experiment |
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| Authors: | Xu Xingang Hu Jianhe Zhang Yanming |
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| Institution: | (1College of Animal Science and Technology, Northwest Sci-Tech University of Agriculture and Forestry, Yangling
Shaanxi 712100; 2Department of Animal Science, Henan Institute of Science and Technology, Xinxiang Henan 453003) |
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| Abstract: | The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting full-length cDNA of CSFV Shimen strain E2 gene into pBABE-puro. Both the recombinant retroviral vector and pVSVg plasmid were transfected into eukaryotic cells 293GP by calcium phosphate transfection method thus the pseudovirus were producesd . The pseudovirus infected eukaryotic cells SP2/0 and expression E2 protein was determined by puromycin-resistant and FACS analysis. BALB/c mice were intraperitoneal injected with SP2/0 cells which expressed CSFV E2 protein. The result showed anti-CSFV E2 antibody was screened by FACS. |
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| Keywords: | CSFV E2 envelope protein Retroviral vector Pseudovirus FACS |
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