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Generation of recombinant vaccinia virus and analysis of virus-induced cell death

STAR Protoc. 2021-10; 
Zhijun Liu, Kidong Kang, Francis Ka-Ming Chan
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Gene Synthesis … using the linearized vector from step 2 and the PCR fragment from step 3. Refer to the manufacturer's instructions (https://www.neb.com/protocols/2014/11/26/nebuilder-hifi-dna- assembly-reaction-protocol) for details on the?… 4%–20% SurePAGE Bis-Tris, GenScript, Cat#?… Get A Quote

摘要

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 Liu et?al. (2021).

關鍵詞

Cell Biology, Immunology, Microbiology, Molecular Biology, Protein expression and purification
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