| 伪狂犬病病毒鄂A株gG^—／LacZ^＋突变株的构建 |
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| 引用本文： | 周复春,陈焕春,方六荣,周锐,吴斌,何启盖.伪狂犬病病毒鄂A株gG^—／LacZ^＋突变株的构建[J].畜牧兽医学报,2001,32(2):134-138. |
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| 作者姓名： | 周复春 陈焕春 方六荣 周锐 吴斌 何启盖 |
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| 基金项目： | “九五”国家科技攻关计划生物技术资助!项目 (96 -C0 1- 0 4- 0 3) |
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| 摘 要： | 以从湖北某猪场分离鉴定的鄂A株为亲本，提取其基因组DNA，克隆含gG基因的SphI/KpnI片段，然后将LacZ基因融合到gG启动子下游，得到重组质粒pUSKZ,将重组质粒与鄂A株基因组共转染PK－15细胞，待细胞完全病变后，在X－gal存在下，作蓝斑筛选纯化。经斑点杂交和PCR扩增证实得到的是基因型为gG^-/LacZ^ 的重组伪狂犬病病毒。
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| 关 键 词： | 伪狂犬病病毒 鄂A株 突变株 |
CONSTRUCTION OF THE MUTANTS OF PSEUDORABIES VIRUS STRAIN Ea WITH THE GENOTYPE gG~-/LacZ~ |
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| Zhou Fuchun,Chen Huanchun,Fang Liurong,Zhou Rui,Wu Bin,He Qigai.CONSTRUCTION OF THE MUTANTS OF PSEUDORABIES VIRUS STRAIN Ea WITH THE GENOTYPE gG~-/LacZ~[J].Acta Veterinaria et Zootechnica Sinica,2001,32(2):134-138. |
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| Authors: | Zhou Fuchun Chen Huanchun Fang Liurong Zhou Rui Wu Bin He Qigai |
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| Abstract: | Pseudorabies virus （PRV） is the causative agent of Aujeszky′s disease in swine．After the bacterial β-galactoside gene was fused to the downstream of the gG promoter，this construct and the genome of pseudorabies virus strain Ea were cotransfected into PK-15．Transfection progeny were plated onto PK-15 and incubated for 2 days under methlcellulose．Then the overlay was removed and replaced by 1％ low melting point agorase in DMEM supplemented with 150ug／ml X-gal．After 2 days，blue plaques were picked and purified 4 times．By means of dot hybridization and PCR β-galactoside gene insered into the genome of pseudorabies virus was demonstrated． |
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| Keywords: | Pseudorabies virus Ea gG-/LacZ Mutant |
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