| 猪伪狂犬病毒浙江株感染性细菌人工染色体克隆的构建 |
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| 引用本文： | 尹文玲,尹龙勃,叶伟成,孙学强,姚火春,张淼涛,王一成,张存.猪伪狂犬病毒浙江株感染性细菌人工染色体克隆的构建[J].病毒学报,2010,26(4). |
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| 作者姓名： | 尹文玲 尹龙勃 叶伟成 孙学强 姚火春 张淼涛 王一成 张存 |
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| 作者单位： | 西北农林科技大学,动物医学院,杨凌,712100;浙江省农业科学院,畜牧兽医研究所,杭州,310021;浙江省农业科学院,畜牧兽医研究所,杭州,310021;南京农业大学,动物医学院,南京,210095;西北农林科技大学,动物医学院,杨凌,712100 |
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| 基金项目： | 公益性行业(农业)科研专项(200803016); 浙江省重大科技专项(优先主题)农业项目(2008C12049) |
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| 摘 要： | 通过同源重组将携带细菌人工染色体(Bacterial Artificial Chromosome,BAC)载体和GFP表达框的pHA2质粒序列插入到PRV病毒的UL23(TK)基因内,获得了重组病毒rPRV-HA2;将该重组病毒的环状基因组电转化到感受态细胞EscherichiacoliDH10B,筛选到病毒的感染性克隆PRV BAC(pPRV)。pPRV转染VeroE6细胞可以重新启动病毒的生产性感染,该拯救病毒的细胞病变和体外增殖特性与rPRV-HA2一致。病毒生长曲线表明TK基因的部分删除和BAC载体的插入不会影响病毒在体外的复制。PRV感染性BAC克隆的成功构建,将方便在大肠杆菌内对病毒基因组进行快速、准确的操作,为进一步开展PRV基因功能和病毒载体研究奠定基础。
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| 关 键 词： | 伪狂犬病毒 细菌人工染色体 感染性克隆 重组病毒 |
Construction of an Infectious Clone of Pseudorabies Virus Strain ZJ Genome Maintained as a Bacterial Artificial Chromosome |
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| YIN Wen-ling,YIN Long-bo,YE Wei-cheng,SUN Xue-qiang,YAO Huo-chun,ZHANG Miao-tao,WANG Yi-cheng,ZHANG Cun.Construction of an Infectious Clone of Pseudorabies Virus Strain ZJ Genome Maintained as a Bacterial Artificial Chromosome[J].Chinese Journal of Virology,2010,26(4). |
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| Authors: | YIN Wen-ling YIN Long-bo YE Wei-cheng SUN Xue-qiang YAO Huo-chun ZHANG Miao-tao WANG Yi-cheng ZHANG Cun |
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| Institution: | YIN Wen-ling1,2,YIN Long-bo1,YE Wei-cheng2,SUN Xue-qiang3,YAO Huo-chun3,ZHANG Miao-tao1,WANG Yi-cheng2,ZHANG Cun2(1.College of Veterinary Medicine,Northwest Agriculture & Forestry University,Yangling 712100,China,2.Institute of Animal Husbandry and Veterinary Science,Zhejiang Academy of Agriculture Sciences,Hangzhou 310021,3.Veterinary Medicine College,Nanjing Agricultural University,Nanjing 210095,China) |
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| Abstract: | pHA2 plasmid sequence,with Bacterial Artificial Chromosome(BAC)vector and the GFP expression cassette,was introduced into the UL23(TK)gene of Pseudorabies virus(PRV)strain ZJ by homologous recombination,and the recombinant PRV(rPRV-HA2)was confirmed and isolated by plaque purification.The circular genome of rPRV-HA2 was electroporated into Escherichia coli strain DH10B and then the PRV BAC(pPRV)was recovered.The transfection of pPRV into VeroE6 cells resulted in productive infection.The rescued virus isolat... |
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| Keywords: | pseudorabies virus bacterial artificial chromosome infectious clone recombinant virus |
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