Protocol detail

Characterization of SARS-CoV-2 M Protein Interactions

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The SARS-CoV M protein (NC_004718), SARS-CoV-2 M protein of IPBCAMS-WH-01/2019 strain (no. EPI_ISL_402123), TBK1 genes and their truncations were cloned into vector VR1012 with the Flag-tag or GST-tag (Sangon Biotech, Shanghai, China). The expression vectors of Flag-Ubi, Flag-K48-Ubi, Flag-K63-Ubi, pIFNβ-Luc, ISRE-luc and Renilla luc were constructed in the previous study (17 (link), 18 (link)). The expression plasmids for IRF3, TANK, IKKϵ, RIG-I and TRAF3 were purchased from PPL Biotech, Jiangsu, China. The expression plasmid for TBK1 and MDA5 was purchased from Miaoling Biotech, Wuhan, China.

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Sui L., Zhao Y., Wang W., Wu P., Wang Z., Yu Y., Hou Z., Tan G, & Liu Q. (2021). SARS-CoV-2 Membrane Protein Inhibits Type I Interferon Production Through Ubiquitin-Mediated Degradation of TBK1. Frontiers in Immunology, 12, 662989.

Publication 2021

Flag-tag GST-tag

Genes Irf3 Mda5 Plasmid Renilla Rig i Sars cov 2 m protein Sars cov m protein Strain Tbk1 Traf3 Vectors

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Variable analysis

independent variables

SARS-CoV M protein (NC_004718)
SARS-CoV-2 M protein of IPBCAMS-WH-01/2019 strain (no. EPI_ISL_402123)
TBK1 genes and their truncations
Flag-Ubi
Flag-K48-Ubi
Flag-K63-Ubi
IKKϵ
RIG-I
TRAF3

dependent variables

Measurement of outcomes from the expression of the above variables

control variables

Vector VR1012 with the Flag-tag or GST-tag
PIFNβ-Luc
ISRE-luc
Renilla luc

positive controls

Expression plasmids for IRF3, TANK, IKKϵ, RIG-I, TRAF3, TBK1, MDA5

negative controls

Vector VR1012 with the Flag-tag or GST-tag
PIFNβ-Luc
ISRE-luc
Renilla luc

Annotations

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