{"title":"委内瑞拉链霉菌中fbiC的缺失使青色荧光蛋白激发-发射范围内的自身荧光消失。","authors":"Parminder Singh Mavi, Klas Flärdh","doi":"10.1099/mic.0.001552","DOIUrl":null,"url":null,"abstract":"<p><p>Autofluorescence poses an impediment to fluorescence microscopy of biological samples. In the Gram-positive, soil-dwelling bacteria of the genus <i>Streptomyces</i>, sources of autofluorescence have not been examined systematically to date. Here, we show that the model organism for the genus, <i>Streptomyces venezuelae</i>, shows autofluorescence in two of the commonly used fluorescence channels for visualizing cyan and green/yellow fluorescent proteins. We identify the source of autofluorescence in the cyan fluorescence channel as redox cofactor factor 420 (F<sub>420</sub>) and target its synthesis to remove it. By deleting the <i>vnz15170</i> (<i>fbiC</i>) gene, which is a key biosynthetic gene for the production of F<sub>420</sub>, we were able to create an autofluorescence-free strain in the cyan range of fluorescence excitation-emission. We demonstrate the usefulness of this strain by imaging the mTurquoise-tagged polar growth-related protein DivIVA and the cell division-related protein FtsZ in the <i>fbiC</i> deletion background. Using live-cell imaging to follow the dynamics of DivIVA and FtsZ, we demonstrate an improved signal-to-noise ratio in the mutant strain. We show that this strain can be a suitable tool for visualizing the localization of proteins in <i>Streptomyces</i> spp. and can facilitate the utilization of multi-colour imaging and fluorescence resonance energy transfer-based imaging.</p>","PeriodicalId":49819,"journal":{"name":"Microbiology-Sgm","volume":"171 4","pages":""},"PeriodicalIF":2.6000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Deletion of <i>fbiC</i> in <i>Streptomyces venezuelae</i> removes autofluorescence in the excitation-emission range of cyan fluorescent protein.\",\"authors\":\"Parminder Singh Mavi, Klas Flärdh\",\"doi\":\"10.1099/mic.0.001552\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Autofluorescence poses an impediment to fluorescence microscopy of biological samples. In the Gram-positive, soil-dwelling bacteria of the genus <i>Streptomyces</i>, sources of autofluorescence have not been examined systematically to date. Here, we show that the model organism for the genus, <i>Streptomyces venezuelae</i>, shows autofluorescence in two of the commonly used fluorescence channels for visualizing cyan and green/yellow fluorescent proteins. We identify the source of autofluorescence in the cyan fluorescence channel as redox cofactor factor 420 (F<sub>420</sub>) and target its synthesis to remove it. By deleting the <i>vnz15170</i> (<i>fbiC</i>) gene, which is a key biosynthetic gene for the production of F<sub>420</sub>, we were able to create an autofluorescence-free strain in the cyan range of fluorescence excitation-emission. We demonstrate the usefulness of this strain by imaging the mTurquoise-tagged polar growth-related protein DivIVA and the cell division-related protein FtsZ in the <i>fbiC</i> deletion background. Using live-cell imaging to follow the dynamics of DivIVA and FtsZ, we demonstrate an improved signal-to-noise ratio in the mutant strain. We show that this strain can be a suitable tool for visualizing the localization of proteins in <i>Streptomyces</i> spp. and can facilitate the utilization of multi-colour imaging and fluorescence resonance energy transfer-based imaging.</p>\",\"PeriodicalId\":49819,\"journal\":{\"name\":\"Microbiology-Sgm\",\"volume\":\"171 4\",\"pages\":\"\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2025-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbiology-Sgm\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1099/mic.0.001552\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbiology-Sgm","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1099/mic.0.001552","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Deletion of fbiC in Streptomyces venezuelae removes autofluorescence in the excitation-emission range of cyan fluorescent protein.
Autofluorescence poses an impediment to fluorescence microscopy of biological samples. In the Gram-positive, soil-dwelling bacteria of the genus Streptomyces, sources of autofluorescence have not been examined systematically to date. Here, we show that the model organism for the genus, Streptomyces venezuelae, shows autofluorescence in two of the commonly used fluorescence channels for visualizing cyan and green/yellow fluorescent proteins. We identify the source of autofluorescence in the cyan fluorescence channel as redox cofactor factor 420 (F420) and target its synthesis to remove it. By deleting the vnz15170 (fbiC) gene, which is a key biosynthetic gene for the production of F420, we were able to create an autofluorescence-free strain in the cyan range of fluorescence excitation-emission. We demonstrate the usefulness of this strain by imaging the mTurquoise-tagged polar growth-related protein DivIVA and the cell division-related protein FtsZ in the fbiC deletion background. Using live-cell imaging to follow the dynamics of DivIVA and FtsZ, we demonstrate an improved signal-to-noise ratio in the mutant strain. We show that this strain can be a suitable tool for visualizing the localization of proteins in Streptomyces spp. and can facilitate the utilization of multi-colour imaging and fluorescence resonance energy transfer-based imaging.
期刊介绍:
We publish high-quality original research on bacteria, fungi, protists, archaea, algae, parasites and other microscopic life forms.
Topics include but are not limited to:
Antimicrobials and antimicrobial resistance
Bacteriology and parasitology
Biochemistry and biophysics
Biofilms and biological systems
Biotechnology and bioremediation
Cell biology and signalling
Chemical biology
Cross-disciplinary work
Ecology and environmental microbiology
Food microbiology
Genetics
Host–microbe interactions
Microbial methods and techniques
Microscopy and imaging
Omics, including genomics, proteomics and metabolomics
Physiology and metabolism
Systems biology and synthetic biology
The microbiome.