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. 2021 Aug 16;20:158. doi: 10.1186/s12934-021-01653-9

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Knockout of the kstd and ksh genes from the HGMS2 strain. a A homologous recombination vector was constructed based on p2NIL plasmid. U: the upstream sequence; D: the downstream sequence. b PCR confirmation of putative kstd-knockout colonies, in comparison with that for the WT strain. M: DNA marker; WT: PCR products amplified from the HGMS2; ΔKstD211: two putative kstd-default mutants. c–e PCR confirmation of putative ksh-knockout colonies, in comparison with that for the WT strain. M: DNA marker; ΔKshA226: a putative HGMS2^{ΔkstD211 + ΔkshA226} mutant; KOA226-Pl: the plasmid p2NIL-Sac-ΔKshA226; WT: the HGMS2 strain; ΔKshA395: a putative HGMS2^{Δkstd211 + ΔkshA395} mutant; KOA395-Pl: the plasmid p2NIL-Sac-ΔKshA395; ΔKshB122a and ΔKshB122: two putative HGMS2^{Δkstd211 + ΔkshB122} mutants; KOB122-Pl: the plasmid p2NIL-Sac-ΔKshB122