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Proteins, Expression, Isolation and Analysis | After centrifugation at 100 g for 10 min at 4 °C, the supernatant was mixed with incubated at 4 °C for 2 h. After washing, the IP products were boiled at 100 °C for 10 min. Samples were separated by 10% Bis-Tris PAGE Gel (GenScript, Nanjing, China) and subjected to immunoblot analysis. The ubiquitin modifications were detected with the anti-Myc (1:10 000, Abmart, M20002) and anti-Ubi P4D1 (1:1000, Santa Cruz, sc-8017) antibodies. The immunoprecipitated OsWRKY proteins were detected with the anti-FLAG (1:10 000, GenScript, A00187) antibody . | Get A Quote |
Plants have intricate mechanisms that tailor their defence responses to pathogens. WRKY transcription factors play a pivotal role in plant immunity by regulating various defence signalling pathways. Many WRKY genes are transcriptionally activated upon pathogen attack, but how their functions are regulated after transcription remains elusive. Here, we show that OsWRKY7 functions as a crucial positive regulator of rice basal immunity against Xanthomonas oryzae pv. oryzae (Xoo). The activity of OsWRKY7 was regulated at both translational and post-translational levels. Two translational products of OsWRKY7 were generated by alternative initiation. The full-length OsWRKY7 protein is normally degraded by the ubiquiti... More