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URL: https://dx.doi.org/10.17504/protocols.io.bdyei7te

Authors: Kizhakke Mattada Sathyan, Thomas G. Scott, Michael J. Guertin

Summary: The next step in developing the ARF-AID system is to tag the gene of interest with AID. The ARF-AID system requires full-length AID (Figure 1) because the characterized interaction domains of AID with ARF are domains III and IV. Domains I and II are involved in the interaction with TIR1. The mini-AID lacks domains III and IV and will not interact with ARF to stabilize the protein in the absence of auxin (Sathyan et al., 2019). Note that the antibiotic selection marker (HygroR) is co-transcribed with AID and the protein products are separated during translation. Therefore, the resistance marker will be expressed at levels comparable to the target protein.

Affiliations: Biochemistry and Molecular Genetics Department, University of Virginia, Biochemistry and Molecular Genetics Department, University of Virginia, Biochemistry and Molecular Genetics Department, University of Virginia ; Center for Public Health Genomics, University of Virginia; Cancer Center, University of Virginia

Version: 1

Publication Date: 2020

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