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. 2020 Dec 3;10:21109. doi: 10.1038/s41598-020-77987-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Pseudo-revertants from the porA deletion mutant. (A) Mutations in porY of the pseudo-revertants. Mutations causing amino acid substitutions were found in the cytoplasmic domain-encoding region of porY. HK, histidine kinase domain; SS, signal sequence; TM, transmembrane region. Circle, square, and triangle indicate amino acid substitutions at 204, 241, and 266, respectively. (B) Colony pigmentation of the wild type, ΔporA, ΔporA/porY(S266W), and ΔporA ΔporX/porY(S266W) strains on the blood agar plate for 6 days. (C) qRT-PCR expression analysis of various T9SS-related genes in the wild type, ΔporA, ΔporA/porA⁺ and ΔporA/porY(S266W) strains. The mean of expression of each wild type gene was regarded as 1. (D) Gingipain activities of the wild type, ΔporA, ΔporA/porY(S266W), and ΔporA ΔporX/porY(S266W) strains. (E) Expression of the porK, porT, porA, porX, and sigP genes in the wild type, ΔporA, ΔporA/porY(S266W), and ΔporA ΔporX/porY(S266W) strains. RNA samples of the strains were subjected to qRT-PCR analysis. The mean expression of each wild type gene was regarded as 1.