| 基于甲病毒复制子的犬瘟热核酸疫苗的构建及表达性研究 |
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| 引用本文： | 孙建涛,侯绍华,于萍,朱鸿飞,梁欠,郝志明. 基于甲病毒复制子的犬瘟热核酸疫苗的构建及表达性研究[J]. 中国畜牧兽医, 2013, 40(9): 96-100 |
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| 作者姓名： | 孙建涛 侯绍华 于萍 朱鸿飞 梁欠 郝志明 |
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| 作者单位： | 1. 天津农学院动物科学系, 天津 300384;2. 中国农业科学院北京畜牧兽医研究所, 北京 100193;3. 中国农业科学院兰州畜牧与兽药研究所, 甘肃兰州 730050 |
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| 基金项目： | 农业部动物疫情监测与防治项目;863动物疫病分子诊断技术研究与产品创制项目(2012AA101302);国家科技支撑计划重点项目(2010BAD04B02);公益性行业(农业)科研专项经费项目(200903027)。 |
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| 摘 要： | 本试验利用T-A克隆技术,构建克隆载体pEASY-Blunt-F,BamHⅠ酶切pEASY-Blunt-F,回收F基因,将其克隆至pSCA1真核表达载体中,获得重组质粒pSCA1-F。经DNA测序、限制性内切酶分析和PCR鉴定,结果表明重组质粒pSCA1-F成功构建。通过间接免疫荧光试验和Western blotting证实F基因在转染细胞中表达。此外,细胞凋亡的检测结果表明,pSCA1-F能引起转染的细胞发生凋亡。
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| 关 键 词： | 核酸疫苗 犬瘟热病毒 F基因 甲病毒复制子 细胞凋亡 |
| 收稿时间： | 2013-01-31 |
Study on the Construction and Expression Characteristics of Canine Distemper DNA Vaccine Based on SFV-replicon |
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| SUN Jian-tao,HOU Shao-hua,YU Ping,ZHU Hong-fei,LIANG Qian,HAO Zhi-ming. Study on the Construction and Expression Characteristics of Canine Distemper DNA Vaccine Based on SFV-replicon[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(9): 96-100 |
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| Authors: | SUN Jian-tao HOU Shao-hua YU Ping ZHU Hong-fei LIANG Qian HAO Zhi-ming |
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| Affiliation: | 1. Department of Animal Science, Tianjin Agricultural University, Tianjin 300384, China;2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;3. Lanzhou Institute of Husbandry and Pharmaceutical Science, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China |
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| Abstract: | The clone vector pEASY-Blunt-F with fusion gene of canine distemper virus(CDV) isolated from vaccine strain was successfully constructed by T-A clone technique and was digested with restriction enzyme of BamHⅠ. The F gene was cloned into eukaryotic expression vector pSCA1. Finally a recombinant plasmid named pSCA1-F was constructed and identified by PCR, restriction enzyme analysis and sequencing. pSCA1-F was transfected into BHK-21 cells by using X-tremeGENE 9 DNA Transfection Reagent,IFA and Western blotting were performed to detect F expressions,the results showed that F gene had expressed in the transfected cells.Furthermore,apoptosis assay confirmed that pSCA1-F could induce the transfected cells apoptosis. |
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| Keywords: | DNA vaccine canine distemper virus F gene SFV-replicon apoptosis |
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