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. doi: 10.1016/j.virs.2022.03.005
Citation: Lifang Song, Bopei Cui, Jinghuan Yang, Xiaotian Hao, Xujia Yan, Jialu Zhang, Dong Liu, Ziyang Song, Qian Wang, Qunying Mao, Zhenglun Liang. Construction and verification of an infectious cDNA clone of coxsackievirus B5 [J].VIROLOGICA SINICA, 2022, 37(3) : 469-471.  http://dx.doi.org/10.1016/j.virs.2022.03.005

柯萨奇病毒B5 cDNA感染性克隆的构建与验证

  • 目的 构建柯萨奇病毒B组5型(Coxsackievirus B5,CV-B5) Faulkner株cDNA全长感染性克隆。方法 根据CV-B5 Faulkner株基因组序列设计特异性引物,将全长cDNA分两段进行PCR扩增,用融合克隆的方法将两个片段无缝连接到pSVA载体上,经转化后获得单克隆菌落后提取质粒,将质粒与T7质粒共转染到HEK-293T细胞,获得拯救后的病毒,并在Vero细胞上传代。通过细胞病变效应(Cytopathic effect,CPE)、病毒滴度测定、生长动力学实验、3日龄BALB/c乳鼠攻毒模型进行毒力评价等对拯救后病毒的生物学特性进行鉴定,并利用二代测序对拯救病毒连续传代的基因组稳定性进行研究。结果 质粒酶切鉴定及测序表明cDNA感染性克隆构建成功。拯救病毒测序结果与母本毒株一致,获得的拯救病毒在Vero细胞的CPE效应、生长特性、以及乳鼠致病力和组织器官分布等生物学特性方面与母本病毒相同,且遗传稳定性好。结论 成功构建CB5感染性克隆并获得拯救病毒,为病毒致病性、毒力位点、动物模型以及疫苗评价等提供一定基础。

Construction and verification of an infectious cDNA clone of coxsackievirus B5

  • Highlights:
    · An infectious cDNA clone of CV-B5 was constructed.
    · The rescued and parental virus possessed similar biological characteristics.
    · The virulence of the rescued virus was similiar to that of the parental virus.
    · Viral distribution and tissue tropism of those two viruses were in agreement.

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    Construction and verification of an infectious cDNA clone of coxsackievirus B5

      Corresponding author: Qunying Mao, maoqunying@126.com
      Corresponding author: Zhenglun Liang, lzhenglun@126.com
    • a National Institute for Food and Drug Control, Beijing 102629, China;

    Abstract: Highlights:
    · An infectious cDNA clone of CV-B5 was constructed.
    · The rescued and parental virus possessed similar biological characteristics.
    · The virulence of the rescued virus was similiar to that of the parental virus.
    · Viral distribution and tissue tropism of those two viruses were in agreement.

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