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Complementation assay.

ZL Zhongyi Lu
TF Ting Fu
TL Tianyi Li
YL Yang Liu
SZ Siyu Zhang
JL Jinquan Li
JD Junbiao Dai
EK Eugene V. Koonin
GL Guohui Li
HC Huiying Chu
ML Meng Li
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The Vps4 coding sequences belonging to Lokiarchaeum_GC14_75, Thorarchaeota_AB_25, Heimdallarchaeota_LC_3, and Odinarchaeota_LCB_4, and CdvC coding sequences belonging to Sulfolobus solfataricus_P2, and Bathyarchaeota (NCBI:protein accession numbers KKK42121.1, OLS30569.1, OLS27542.1, OLS18192.1, AAK41192.1, and WP_119819537.1, respectively) were codon optimized by GeneDesign (http://54.235.254.95/gd/) for expression in S. cerevisiae, before synthesis by BGI Genomics Co., Ltd. (54). To eliminate the interference of transcriptional level factors, a native promoter region of S. cerevisiae BY4741 vps4 (a 500-bp DNA sequence region upstream from the ATG start codon of this gene) was used to drive the coding sequences. Then, we assembled the coding sequences, the S. cerevisiae vps4 native promoter, and an S. cerevisiae CYC1 (cytochrome c isoform 1) terminator into a pPOT-RFP vector according to a developed YeastFab Assembly protocol (31). In addition, the pPOT-RFP vector containing the entire S. cerevisiae BY4741 vps4 with its native promoter and the CYC1 terminator were transformed into the S. cerevisiae vps4 null mutant (32), and this reconstituted strain was designated the “+S. cerevisiae” strain shown in the figures. In this study, both the S. cerevisiae and S. cerevisiae vps4Δ were transformed with the pPOT-RFP vector as the control.

Copyright and License information:  ©2020 Lu et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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