Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria

Tian, Di; Liu, Yiyuan; Zhang, Yueyue; Liu, Yunfei; Xia, Yang; Xu, Boying; Xu, Jian; Yomo, Tetsuya

doi:10.3390/bioengineering11010081

Open AccessArticle

Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria

by

Di Tian

,

Yiyuan Liu

,

Yueyue Zhang

,

Yunfei Liu

,

Yang Xia

,

Boying Xu

,

Jian Xu

^*

and

Tetsuya Yomo

^*

Laboratory of Biology and Information Science, School of Life Sciences, East China Normal University, Shanghai 200062, China

^*

Authors to whom correspondence should be addressed.

Bioengineering 2024, 11(1), 81; https://doi.org/10.3390/bioengineering11010081

Submission received: 19 December 2023 / Revised: 9 January 2024 / Accepted: 12 January 2024 / Published: 15 January 2024

(This article belongs to the Section Cellular and Molecular Bioengineering)

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Abstract

Cell-wall-less (L-form) bacteria exhibit morphological complexity and heterogeneity, complicating quantitative analysis of them under internal and external stimuli. Stable and efficient labeling is needed for the fluorescence-based quantitative cell analysis of L-forms during growth and proliferation. Here, we evaluated the expression of multiple fluorescent proteins (FPs) under different promoters in the Bacillus subtilis L-form strain LR2 using confocal microscopy and imaging flow cytometry. Among others, P_ylb-derived NBP3510 showed a superior performance for inducing several FPs including EGFP and mKO2 in both the wild-type and L-form strains. Moreover, NBP3510 was also active in Escherichia coli and its L-form strain NC-7. Employing these established FP-labeled strains, we demonstrated distinct morphologies in the L-form bacteria in a quantitative manner. Given cell-wall-deficient bacteria are considered protocell and synthetic cell models, the generated cell lines in our work could be valuable for L-form-based research.

Keywords: cell-wall-less bacteria; L-form; heterogeneity; quantification analysis; fluorescent protein labeling

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MDPI and ACS Style

Tian, D.; Liu, Y.; Zhang, Y.; Liu, Y.; Xia, Y.; Xu, B.; Xu, J.; Yomo, T. Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria. Bioengineering 2024, 11, 81. https://doi.org/10.3390/bioengineering11010081

AMA Style

Tian D, Liu Y, Zhang Y, Liu Y, Xia Y, Xu B, Xu J, Yomo T. Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria. Bioengineering. 2024; 11(1):81. https://doi.org/10.3390/bioengineering11010081

Chicago/Turabian Style

Tian, Di, Yiyuan Liu, Yueyue Zhang, Yunfei Liu, Yang Xia, Boying Xu, Jian Xu, and Tetsuya Yomo. 2024. "Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria" Bioengineering 11, no. 1: 81. https://doi.org/10.3390/bioengineering11010081

APA Style

Tian, D., Liu, Y., Zhang, Y., Liu, Y., Xia, Y., Xu, B., Xu, J., & Yomo, T. (2024). Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria. Bioengineering, 11(1), 81. https://doi.org/10.3390/bioengineering11010081

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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Implementation of Fluorescent-Protein-Based Quantification Analysis in L-Form Bacteria

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