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  • Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta.

Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta.

Bio-protocol (2018-05-05)
Lijia Zheng, Chao Zhang, Jianguo Wu, Yi Li
초록

MicroRNAs (miRNAs) play important roles in plant growth, development, and response to infection by microbes. Double-stranded RNA binding protein 1 (DRB1) facilitates the processing of primary miRNA transcripts into mature miRNAs. Recently, we found that NS3 protein encoded by rice stripe virus (RSV) associates with DRB1 and promotes miRNA biogenesis during RSV infection ( Zheng et al., 2017 ). RNA co-immunoprecipitation (RIP) method was applied to identity association patterns among DRB1, NS3, and miRNA transcript.

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Sigma-Aldrich
TERGITOL solution, Type NP-40, 70% in H2O
Sigma-Aldrich
Formaldehyde solution, for molecular biology, 36.5-38% in H2O
Sigma-Aldrich
Anti-c-Myc antibody, Mouse monoclonal, clone 9E10, purified from hybridoma cell culture
Sigma-Aldrich
Triton X-100, BioXtra
Sigma-Aldrich
Magnesium chloride, anhydrous, ≥98%
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, for molecular biology