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. 2017 May 11;8:853. doi: 10.3389/fmicb.2017.00853

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Copyright © 2017 Chen, Xu, Tao, Li, Nan, Wang, Tian and Chen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Identification of the interaction of NSP7α with NSP9. (A) Yeast two-hybrid assays. The yeast cells were co-transformed with pGADT7-nsp7α and pGBKT7-nsp9 constructs, subjected to 10-fold serial dilutions and plated on SD/-Ade/-His/-Leu/-Trp medium. The cells co-transformed with pGBK7-P53 and pGADT7-T were used as positive interaction controls, and the group with pGBKT7-Lam and pGADT7-T was used as a negative control. (B) Pull-down assays. FLAG-tagged NSP7α and HIS-tagged NSP9 were expressed in E. coli, mixed together, and the protein mixture was purified with nickel magnetic beads. HIS-tagged and FLAG-tagged proteins pulled down by the beads were, respectively, analyzed by Western blot with antibodies against HIS-tag and FLAG-tag.