Generation of Stable Cell Line Expressing T7 Polymerase

BSR-T7/5 cells stably expressing the T7 phage RNA polymerase were generated by Buchholz et al. [90 (link)], and were a kind gift from Prof. Xiufan Liu (Key Laboratory of Animal Infectious Diseases, Yangzhou University, China). The parental NDV (rSS1GFP) and the mutant NDV (rSS1GFP-M/NLSm) carrying M/NLS mutation were generated in our previous study [10 (link)], which were plaque purified three times in chicken embryonic fibroblasts and propagated once in specific pathogen-free (SPF) embryonated chicken eggs. The rabbit polyclonal antibodies against NDV M protein and NP protein was prepared in our laboratory. The rabbit polyclonal antibodies against ENY2 (DF12979), BRD2 (DF12857), RPL18 (DF3700), RPL34 (DF3708), PSMD3 (DF3645), RAB12 (DF12459), TRAF6 (AF5376), NF-κB p65 (AF5006), phospho-NF-κB p65 (Ser536) (AF2006), GAPDH (AF7021), GFP (T0006), HA (T0050) and IL-2 (AF5105) were purchased from Affinity Biosciences (USA). The rabbit polyclonal antibodies against POP1 (ab254978), SWD2 (ab220240), UBR5 (ab70311), STX5 (ab217130), and rabbit monoclonal antibody against phospho-TIFA (Thr9) (ab214815) were purchased from Abcam (USA). The rabbit polyclonal antibody against TIFA (61358S) was purchased from Cell Signaling Technology (USA).

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Duan Z., Yuan C., Han Y., Zhou L., Zhao J., Ruan Y., Chen J., Ni M, & Ji X. (2020). TMT-based quantitative proteomics analysis reveals the attenuated replication mechanism of Newcastle disease virus caused by nuclear localization signal mutation in viral matrix protein. Virulence, 11(1), 607-635.

Publication 2020

AF5006 ab70311

Animal laboratory Antibodies Antibody Cells Chicken Eggs Embryonic Fibroblasts Gapdh Infectious diseases M protein Monoclonal antibody Mutation Nf κb p65 Parental Plaque Protein Rab12 Rabbit Specific pathogen free T7 phage rna polymerase Traf6

Corresponding Organization : Guizhou University

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Variable analysis

independent variables

RSS1GFP (parental NDV)
RSS1GFP-M/NLSm (mutant NDV carrying M/NLS mutation)

dependent variables

Viral propagation in chicken embryonic fibroblasts and specific pathogen-free (SPF) embryonated chicken eggs

control variables

BSR-T7/5 cells stably expressing the T7 phage RNA polymerase
Chicken embryonic fibroblasts and specific pathogen-free (SPF) embryonated chicken eggs used for viral propagation
Rabbit polyclonal antibodies against NDV M protein, NP protein, ENY2, BRD2, RPL18, RPL34, PSMD3, RAB12, TRAF6, NF-κB p65, phospho-NF-κB p65 (Ser536), GAPDH, GFP, HA, and IL-2
Rabbit polyclonal antibodies against POP1, SWD2, UBR5, STX5, and rabbit monoclonal antibody against phospho-TIFA (Thr9)
Rabbit polyclonal antibody against TIFA

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