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. 2017 Dec 22;8:2577. doi: 10.3389/fmicb.2017.02577

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Copyright © 2017 Zhu, Chen, Li and Wen.

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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Expression analysis of avaR1 and related genes. (A) Transcriptional profile of avaR1 during avermectin production process in WT grown in FM-I. Relative value of avaR1 on day 1 was assigned as 1. Error bars: SD from three replicates. (B) Western blotting analysis of AvaR1 protein expression profile during fermentation process. AvaR1 temporal expression in strain ΔavaR1/avaR1-3FLAG grown in FM-I was analyzed using ANTI-FLAG mAb. 100 μg total protein was added in each lane. Loading control: Coomassie Blue staining of total protein. (C) qRT-PCR analysis of aveR, aveA1, cyp17, aco, and three avaR genes in WT and ΔavaR1 grown in FM-I. Value for each gene was expressed relative to that of WT on day 2, which was assigned as 1. avaR1, 125-bp transcript amplified from the remainder avaR1 ORF in ΔavaR1 with primers ZJY129 and ZJY130. Error bars: SD from three replicates. NS, not significant; ^*, P < 0.05; ^**, P < 0.01; ^***, P < 0.001 (Student's t-test).