Site-directed Mutagenesis of UPF0118

All the site-directed or C terminus-truncated variants of UPF0118 were constructed using pET22b-P-UPF0118 as a template and the corresponding primers listed in Supplementary Table S1 via a Fast Mutagenesis System kit purchased from TransGen Biotech Co., Ltd. (Beijing, China), as described in our recent studies (Shao et al., 2018 (link); Xu et al., 2019 (link)). All the final UPF0118 variants were re-sequenced to confirm the accuracy of mutagenesis, and the corresponding plasmids were transformed into E. coli KNabc for growth tests and Na⁺(Li⁺)/H⁺ antiport activity assays.

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Wang L., Zou Q., Yan M., Wang Y., Guo S., Zhang R., Song Y., Li X., Chen H., Shao L., Meng L, & Jiang J. (2020). Polar or Charged Residues Located in Four Highly Conserved Motifs Play a Vital Role in the Function or pH Response of a UPF0118 Family Na+(Li+)/H+ Antiporter. Frontiers in Microbiology, 11, 841.

Publication 2020

Fast Mutagenesis System kit

Antiport Assays E coli Mutagenesis Plasmids Primers

Corresponding Organization : Northeast Agricultural University

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Variable analysis

independent variables

Site-directed or C terminus-truncated variants of UPF0118

dependent variables

Na+(Li+)/H+ antiport activity

control variables

PET22b-P-UPF0118 as a template
Corresponding primers listed in Supplementary Table S1
Fast Mutagenesis System kit from TransGen Biotech Co., Ltd.
E. coli KNabc for growth tests and Na+(Li+)/H+ antiport activity assays

positive controls

None specified

negative controls

None specified

Annotations

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