Generation of recombinant vaccinia virus and analysis of virus-induced cell death.
Generation of recombinant vaccinia virus and analysis of virus-induced cell death.
复制标题
DOI：
10.1016/j.xpro.2021.100871
复制
发表时间：
2021-12-17
期刊：
影响因子：
--
通讯作者：
Ka-Ming Chan F
中科院分区：
文献类型：
--
作者：
Liu Z;Kang K;Ka-Ming Chan F
牛痘病毒是一种大的双链DNA病毒，广泛用于表达来自不同来源的外源基因。我们产生了表达病毒抑制剂的重组牛痘病毒，以检查其对病毒诱导的坏死性凋亡的影响。我们提供了一个详细的协议来描述重组牛痘病毒的产生，蛋白质表达的验证，并使用活细胞成像确定坏死性凋亡。该方法可适用于检查其他细胞死亡调节剂对病毒诱导的细胞死亡的影响。有关本方案使用和执行的完整详细信息，请参阅。构建用于产生重组牛痘病毒的靶向载体基于噬斑大小的重组牛痘病毒的选择重组牛痘病毒的扩增验证靶蛋白表达并检查对病毒诱导的细胞死亡的影响牛痘病毒是一种大的双链DNA病毒，其被广泛用于表达来自不同来源的外源基因。我们产生了表达病毒抑制剂的重组牛痘病毒，以检查其对病毒诱导的坏死性凋亡的影响。我们提供了一个详细的协议来描述重组牛痘病毒的产生，蛋白质表达的验证，并使用活细胞成像确定坏死性凋亡。该方法可适用于检查其他细胞死亡调节剂对病毒诱导的细胞死亡的影响。
Vaccinia virus is a large double-stranded DNA virus that is widely used to express foreign genes from different origins. We generated recombinant vaccinia virus that expresses a viral inhibitor to examine its effect on virus-induced necroptosis. We provide a detailed protocol to describe the generation of recombinant vaccinia virus, validation of protein expression, and determination of necroptosis using live cell imaging. This approach can be adapted to examine the effect of other cell death regulators on virus-induced cell death. For complete details on the use and execution of this protocol, please refer to. Construction of targeting vectors for generation of recombinant vaccinia virus Selection of recombinant vaccinia virus based on plaque size Amplification of recombinant vaccinia virus Validation of target protein expression and examine the effect on virus-induced cell death Vaccinia virus is a large double-stranded DNA virus that is widely used to express foreign genes from different origins. We generated recombinant vaccinia virus that expresses a viral inhibitor to examine its effect on virus-induced necroptosis. We provide a detailed protocol to describe the generation of recombinant vaccinia virus, validation of protein expression, and determination of necroptosis using live cell imaging. This approach can be adapted to examine the effect of other cell death regulators on virus-induced cell death.
登录
查看更多内容
影响因子：
--
作者：
Wallberg, Fredrik;Tenev, Tencho;Meier, Pascal
通讯作者：
Meier, Pascal
影响因子：
32.4
作者：
Liu Z;Nailwal H;Rector J;Rahman MM;Sam R;McFadden G;Chan FK
通讯作者：
Chan FK
影响因子：
3.5
作者：
BLASCO, R;MOSS, B
通讯作者：
MOSS, B
影响因子：
4
作者：
SUTHERST, RW;WAGLAND, BM;ROBERTS, JA
通讯作者：
ROBERTS, JA