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Vitamin B12 as a source of variability in isotope effects for chloroform biotransformation by Dehalobacter 维生素 B12 是 Dehalobacter 进行氯仿生物转化时同位素效应变化的来源。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-08-27 DOI: 10.1002/mbo3.1433
Elizabeth Phillips, Katherine Picott, Steffen Kümmel, Olivia Bulka, Elizabeth Edwards, Po-Hsiang Wang, Matthias Gehre, Ivonne Nijenhuis, Barbara S. Lollar
{"title":"Vitamin B12 as a source of variability in isotope effects for chloroform biotransformation by Dehalobacter","authors":"Elizabeth Phillips,&nbsp;Katherine Picott,&nbsp;Steffen Kümmel,&nbsp;Olivia Bulka,&nbsp;Elizabeth Edwards,&nbsp;Po-Hsiang Wang,&nbsp;Matthias Gehre,&nbsp;Ivonne Nijenhuis,&nbsp;Barbara S. Lollar","doi":"10.1002/mbo3.1433","DOIUrl":"10.1002/mbo3.1433","url":null,"abstract":"<p>Carbon and chlorine isotope effects for biotransformation of chloroform by different microbes show significant variability. Reductive dehalogenases (RDase) enzymes contain different cobamides, affecting substrate preferences, growth yields, and dechlorination rates and extent. We investigate the role of cobamide type on carbon and chlorine isotopic signals observed during reductive dechlorination of chloroform by the RDase CfrA. Microcosm experiments with two subcultures of a <i>Dehalobacter</i>-containing culture expressing CfrA—one with exogenous cobamide (Vitamin B<sub>12</sub>, B12<sup>+</sup>) and one without (to drive native cobamide production)—resulted in a markedly smaller carbon isotope enrichment factor (<i>ε</i><sub>C, bulk</sub>) for B12<sup>−</sup> (−22.1 ± 1.9‰) compared to B12<sup>+</sup> (−26.8 ± 3.2‰). Both cultures exhibited significant chlorine isotope fractionation, and although a lower <i>ε</i><sub>Cl, bulk</sub> was observed for B12<sup>−</sup> (−6.17 ± 0.72‰) compared to B12<sup>+</sup> (−6.86 ± 0.77‰) cultures, these values are not statistically different. Importantly, dual-isotope plots produced identical slopes of <i>Λ</i><sub>Cl/C</sub> (<i>Λ</i><sub>Cl/C, B12+</sub> = 3.41 ± 0.15, <i>Λ</i><sub>Cl/C, B12</sub>− = 3.39 ± 0.15), suggesting the same reaction mechanism is involved in both experiments, independent of the lower cobamide bases. A nonisotopically fractionating masking effect may explain the smaller fractionations observed for the B12<sup>−</sup> containing culture.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1433","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142073250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Extraction of high-molecular-weight DNA from Streptococcus spp. for nanopore sequencing in resource-limited settings 从链球菌中提取高分子量 DNA,用于资源有限环境下的纳米孔测序。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-08-21 DOI: 10.1002/mbo3.1432
Suvra Das, JérÔme Delamare-Deboutteville, Andrew C. Barnes, Oleksandra Rudenko
{"title":"Extraction of high-molecular-weight DNA from Streptococcus spp. for nanopore sequencing in resource-limited settings","authors":"Suvra Das,&nbsp;JérÔme Delamare-Deboutteville,&nbsp;Andrew C. Barnes,&nbsp;Oleksandra Rudenko","doi":"10.1002/mbo3.1432","DOIUrl":"10.1002/mbo3.1432","url":null,"abstract":"<p>The long-read sequencing platform MinION, developed by Oxford Nanopore Technologies, enables the sequencing of bacterial genomes in resource-limited settings, such as field conditions or low- and middle-income countries. For this purpose, protocols for extracting high-molecular-weight DNA using nonhazardous, inexpensive reagents and equipment are needed, and some methods have been developed for gram-negative bacteria. However, we found that without modification, these protocols are unsuitable for gram-positive <i>Streptococcus</i> spp., a major threat to fish farming and food security in low- and middle-income countries. Multiple approaches were evaluated, and the most effective was an extraction method using lysozyme, sodium dodecyl sulfate, and proteinase K for lysis of bacterial cells and magnetic beads for DNA recovery. We optimized the method to consistently achieve sufficient yields of pure high-molecular-weight DNA with minimal reagents and time and developed a version of the protocol which can be performed without a centrifuge or electrical power. The suitability of the method was verified by MinION sequencing and assembly of 12 genomes of epidemiologically diverse fish-pathogenic <i>Streptococcus iniae</i> and <i>Streptococcus agalactiae</i> isolates. The combination of effective high-molecular-weight DNA extraction and MinION sequencing enabled the discovery of a naturally occurring 15 kb low-copy number mobilizable plasmid in <i>S. iniae</i>, which we name pSI1. We expect that our resource-limited settings-adapted protocol for high-molecular-weight DNA extraction could be implemented successfully for similarly recalcitrant-to-lysis gram-positive bacteria, and it represents a method of choice for MinION-based disease diagnostics in low- and middle-income countries.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1432","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142009034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Differential association of key bacterial groups with diatoms and Phaeocystis spp. during spring blooms in the Southern Ocean 南大洋春季水华期间主要细菌群与硅藻和 Phaeocystis spp.
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-08-09 DOI: 10.1002/mbo3.1428
Nyree J. West, Marine Landa, Ingrid Obernosterer
{"title":"Differential association of key bacterial groups with diatoms and Phaeocystis spp. during spring blooms in the Southern Ocean","authors":"Nyree J. West,&nbsp;Marine Landa,&nbsp;Ingrid Obernosterer","doi":"10.1002/mbo3.1428","DOIUrl":"10.1002/mbo3.1428","url":null,"abstract":"<p>Interactions between phytoplankton and heterotrophic bacteria significantly influence the cycling of organic carbon in the ocean, with many of these interactions occurring at the micrometer scale. We explored potential associations between specific phytoplankton and bacteria in two size fractions, 0.8–3 µm and larger than 3 µm, at three naturally iron-fertilized stations and one high nutrient low chlorophyll station in the Southern Ocean. The composition of phytoplankton and bacterial communities was determined by sequencing the <i>rbcL</i> gene and 16S rRNA gene from DNA and RNA extracts, which represent presence and potential activity, respectively. Diatoms, particularly <i>Thalassiosira</i>, contributed significantly to the DNA sequences in the larger size fractions, while haptophytes were dominant in the smaller size fraction. Correlation analysis between the most abundant phytoplankton and bacterial operational taxonomic units revealed strong correlations between <i>Phaeocystis</i> and picoeukaryotes with SAR11, SAR116, <i>Magnetospira</i>, and <i>Planktomarina</i>. In contrast, most <i>Thalassiosira</i> operational taxonomic units showed the highest correlations with <i>Polaribacter</i>, <i>Sulfitobacteria</i>, <i>Erythrobacter</i>, and <i>Sphingobium</i>, while <i>Fragilariopsis</i>, <i>Haslea</i>, and <i>Thalassionema</i> were correlated with OM60, <i>Fluviicola</i>, and <i>Ulvibacter</i>. Our in-situ observations suggest distinct associations between phytoplankton and bacterial taxa, which could play crucial roles in nutrient cycling in the Southern Ocean.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11310772/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141907053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Exploring the resilience and stability of a defined human gut microbiota consortium: An isothermal microcalorimetric study 探索确定的人类肠道微生物群联合体的恢复力和稳定性:等温微量热法研究
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-08-08 DOI: 10.1002/mbo3.1430
Anna Kattel, Valter Aro, Petri-Jaan Lahtvee, Jekaterina Kazantseva, Arvi Jõers, Ranno Nahku, Isma Belouah
{"title":"Exploring the resilience and stability of a defined human gut microbiota consortium: An isothermal microcalorimetric study","authors":"Anna Kattel,&nbsp;Valter Aro,&nbsp;Petri-Jaan Lahtvee,&nbsp;Jekaterina Kazantseva,&nbsp;Arvi Jõers,&nbsp;Ranno Nahku,&nbsp;Isma Belouah","doi":"10.1002/mbo3.1430","DOIUrl":"10.1002/mbo3.1430","url":null,"abstract":"<p>The gut microbiota significantly contributes to human health and well-being. The aim of this study was to evaluate the stability and resilience of a consortium composed of three next-generation probiotics (NGPs) candidates originally found in the human gut. The growth patterns of <i>Akkermansia muciniphila</i>, <i>Bacteroides thetaiotaomicron</i>, and <i>Faecalibacterium prausnitzii</i> were studied both individually and consortium. The growth kinetics of Akkermansia muciniphila (<i>A. muciniphila</i>), Bacteroides thetaiotaomicron (<i>B. thetaiotaomicron</i>), and Faecalibacterium prausnitzii (<i>F. prausnitzii</i>) were characterized both individually and in consortium using isothermal microcalorimetry and 16S ribosomal RNA next-generation sequencing. The consortium reached stability after three passages and demonstrated resilience to changes in its initial composition. The concentration of butyrate produced was nearly twice as high in the consortium compared to the monoculture of F. prausnitzii. The experimental conditions and methodologies used in this article are a solid foundation for developing further complex consortia.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11307317/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141902302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
New functions of pirin proteins and a 2-ketoglutarate: Ferredoxin oxidoreductase ortholog in Bacteroides fragilis metabolism and their impact on antimicrobial susceptibility to metronidazole and amixicile pirin 蛋白和 2-ketoglutarate:脆弱拟杆菌新陈代谢中铁红蛋白氧化还原酶同源物的新功能及其对甲硝唑和阿米西利抗菌药敏感性的影响。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-08-07 DOI: 10.1002/mbo3.1429
Andrea M. Gough, Anita C. Parker, Patricia J. O'Bryan, Terence R. Whitehead, Sourav Roy, Brandon L. Garcia, Paul S. Hoffman, C. Jeffrey Smith, Edson R. Rocha
{"title":"New functions of pirin proteins and a 2-ketoglutarate: Ferredoxin oxidoreductase ortholog in Bacteroides fragilis metabolism and their impact on antimicrobial susceptibility to metronidazole and amixicile","authors":"Andrea M. Gough,&nbsp;Anita C. Parker,&nbsp;Patricia J. O'Bryan,&nbsp;Terence R. Whitehead,&nbsp;Sourav Roy,&nbsp;Brandon L. Garcia,&nbsp;Paul S. Hoffman,&nbsp;C. Jeffrey Smith,&nbsp;Edson R. Rocha","doi":"10.1002/mbo3.1429","DOIUrl":"10.1002/mbo3.1429","url":null,"abstract":"<p>The understanding of how central metabolism and fermentation pathways regulate antimicrobial susceptibility in the anaerobic pathogen <i>Bacteroides fragilis</i> is still incomplete. Our study reveals that <i>B. fragilis</i> encodes two iron-dependent, redox-sensitive regulatory pirin protein genes, <i>pir1</i> and <i>pir2</i>. The mRNA expression of these genes increases when exposed to oxygen and during growth in iron-limiting conditions. These proteins, Pir1 and Pir2, influence the production of short-chain fatty acids and modify the susceptibility to metronidazole and amixicile, a new inhibitor of pyruvate: ferredoxin oxidoreductase in anaerobes. We have demonstrated that Pir1 and Pir2 interact directly with this oxidoreductase, as confirmed by two-hybrid system assays. Furthermore, structural analysis using AlphaFold2 predicts that Pir1 and Pir2 interact stably with several central metabolism enzymes, including the 2-ketoglutarate:ferredoxin oxidoreductases Kor1AB and Kor2CDAEBG. We used a series of metabolic mutants and electron transport chain inhibitors to demonstrate the extensive impact of bacterial metabolism on metronidazole and amixicile susceptibility. We also show that amixicile is an effective antimicrobial against <i>B. fragilis</i> in an experimental model of intra-abdominal infection. Our investigation led to the discovery that the <i>kor2AEBG</i> genes are essential for growth and have dual functions, including the formation of 2-ketoglutarate via the reverse TCA cycle. However, the metabolic activity that bypasses the function of Kor2AEBG following the addition of phospholipids or fatty acids remains undefined. Overall, our study provides new insights into the central metabolism of <i>B. fragilis</i> and its regulation by pirin proteins, which could be exploited for the development of new narrow-spectrum antimicrobials in the future.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11304471/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141897758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Reassessment of the genetic basis of natural rifampin resistance in the genus Rickettsia 重新评估立克次体属天然利福平抗药性的遗传基础。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-07-31 DOI: 10.1002/mbo3.1431
Julien Amoros, Noor Fattar, Marie Buysse, Meriem Louni, Joanne Bertaux, Didier Bouchon, Olivier Duron
{"title":"Reassessment of the genetic basis of natural rifampin resistance in the genus Rickettsia","authors":"Julien Amoros,&nbsp;Noor Fattar,&nbsp;Marie Buysse,&nbsp;Meriem Louni,&nbsp;Joanne Bertaux,&nbsp;Didier Bouchon,&nbsp;Olivier Duron","doi":"10.1002/mbo3.1431","DOIUrl":"10.1002/mbo3.1431","url":null,"abstract":"<p><i>Rickettsia</i>, a genus of obligate intracellular bacteria, includes species that cause significant human diseases. This study challenges previous claims that the Leucine-973 residue in the RNA polymerase beta subunit is the primary determinant of rifampin resistance in <i>Rickettsia</i>. We investigated a previously untested <i>Rickettsia</i> species, <i>R. lusitaniae</i>, from the Transitional group and found it susceptible to rifampin, despite possessing the Leu-973 residue. Interestingly, we observed the conservation of this residue in several rifampin-susceptible species across most <i>Rickettsia</i> phylogenetic groups. Comparative genomics revealed potential alternative resistance mechanisms, including additional amino acid variants that could hinder rifampin binding and genes that could facilitate rifampin detoxification through efflux pumps. Importantly, the evolutionary history of <i>Rickettsia</i> genomes indicates that the emergence of natural rifampin resistance is phylogenetically constrained within the genus, originating from ancient genetic features shared among a unique set of closely related <i>Rickettsia</i> species. Phylogenetic patterns appear to be the most reliable predictors of natural rifampin resistance, which is confined to a distinct monophyletic subclade known as Massiliae. The distinctive features of the RNA polymerase beta subunit in certain untested <i>Rickettsia</i> species suggest that <i>R. raoultii</i>, <i>R. amblyommatis</i>, <i>R. gravesii</i>, and <i>R. kotlanii</i> may also be naturally rifampin-resistant species.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11289727/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141855996","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
In vivo examination of pathogenicity and virulence in environmentally isolated Vibrio vulnificus 对环境分离的弧菌的致病性和毒力进行体内检测。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-07-23 DOI: 10.1002/mbo3.1427
Shannon E. Pipes, Charles R. Lovell, Katie L. Kathrein
{"title":"In vivo examination of pathogenicity and virulence in environmentally isolated Vibrio vulnificus","authors":"Shannon E. Pipes,&nbsp;Charles R. Lovell,&nbsp;Katie L. Kathrein","doi":"10.1002/mbo3.1427","DOIUrl":"10.1002/mbo3.1427","url":null,"abstract":"<p>Human exposure to <i>Vibrio vulnificus</i>, a gram-negative, halophilic environmental pathogen, is increasing. Despite this, the mechanisms of its pathogenicity and virulence remain largely unknown. Each year, hundreds of infections related to <i>V. vulnificus</i> occur, leading to hospitalization in 92% of cases and a mortality rate of 35%. The infection is severe, typically contracted through the consumption of contaminated food or exposure of an open wound to contaminated water. This can result in necrotizing fasciitis and the need for amputation of the infected tissue. Although several genes (<i>rtxA1, vvpE</i>, and <i>vvhA</i>) have been implicated in the pathogenicity of this organism, a defined mechanism has not been discovered. In this study, we examine environmentally isolated <i>V. vulnificus</i> strains using a zebrafish model (<i>Danio rerio</i>) to investigate their virulence capabilities. We found significant variation in virulence between individual strains. The commonly used marker gene of disease-causing strains, <i>vcgC</i>, did not accurately predict the more virulent strains. Notably, the least virulent strain in the study, <i>V. vulnificus</i> Sept WR1-BW6, which tested positive for <i>vcgC, vvhA</i>, and <i>rtxA1</i>, did not cause severe disease in the fish and was the only strain that did not result in any mortality. Our study demonstrates that virulence varies greatly among different environmental strains and cannot be accurately predicted based solely on genotype.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11264103/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141748591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bacterial diversity and geomicrobiology of Winter Wonderland ice cave, Utah, USA 美国犹他州冬季仙境冰洞的细菌多样性和地质微生物学。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-07-12 DOI: 10.1002/mbo3.1426
Miranda Herschel Seixas, Jeffrey S. Munroe, Erin M. Eggleston
{"title":"Bacterial diversity and geomicrobiology of Winter Wonderland ice cave, Utah, USA","authors":"Miranda Herschel Seixas,&nbsp;Jeffrey S. Munroe,&nbsp;Erin M. Eggleston","doi":"10.1002/mbo3.1426","DOIUrl":"10.1002/mbo3.1426","url":null,"abstract":"<p>The Winter Wonderland ice cave, located at an elevation of 3140 m above sea level in the Uinta Mountains of northern Utah, USA, maintains a constant sub-zero temperature. Seasonal snowmelt and rain enter the cave, freeze on the surface of the existing ice, and contribute to a 3-m-thick layered ice mass. This ice mass contains organic matter and cryogenic cave carbonates (CCCs) that date back centuries. In this study, samples of ice, liquid water, and exposed CCCs were collected to examine the bacterial communities within the cave and to determine if these communities vary spatially and between sample types. Flow cytometry showed that cell counts are an order of magnitude higher in liquid water samples than in ice. Epifluorescence microscopy and scanning electron microscopy imaging revealed potential coccoid and bacillus microbial morphologies in water samples and putative cells or calcite spherules in the CCCs. The diversity of bacteria associated with soil, identified through sequence-based analysis, supports the hypothesis that water enters the cave by filtering through soil and bedrock. A differential abundance of bacterial taxa was observed between sample types, with the greatest diversity found in CCCs. This supports a geomicrobiological framework where microbes aggregate in the water, sink into a concentrated layer, and precipitate out of the ice with the CCCs, thereby reducing the cell counts in the ice. These CCCs may provide essential nutrients for the bacteria or could themselves be products of biomineralization.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11241547/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141590740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A deoxyviolacein-based transposon insertion vector for pigmented tracer studies 基于脱氧紫草素的转座子插入载体,用于色素示踪剂研究。
IF 3.9 3区 生物学
MicrobiologyOpen Pub Date : 2024-07-10 DOI: 10.1002/mbo3.1425
Benjamin R. Dietz, Tyler J. Nelson, Neil E. Olszewski, Brett M. Barney
{"title":"A deoxyviolacein-based transposon insertion vector for pigmented tracer studies","authors":"Benjamin R. Dietz,&nbsp;Tyler J. Nelson,&nbsp;Neil E. Olszewski,&nbsp;Brett M. Barney","doi":"10.1002/mbo3.1425","DOIUrl":"10.1002/mbo3.1425","url":null,"abstract":"<p>Pigments provide a simple means to rapidly visually ascertain the quantities or presence of specific microbes in a complex community. The selection of pigment-producing colonies that are simple to differentiate from common colony phenotypes provides a high degree of certainty for the identity of pigment-tagged strains. Successful employment of pigment production is dependent on various intrinsic factors related to proper levels of gene expression and pigment production that are not always easy to predict and vary within each microbe. We have constructed a simple transposon system that incorporates the genes for the production of deoxyviolacein, a pigment produced from intracellular reserves of the amino acid tryptophan, to randomly insert these genes throughout the genome. This tool allows the user to select from many thousands of potential sites throughout a bacterial genome for an ideal location to generate the desired amount of pigment. We have applied this system to a small selection of endophytes and other model bacteria to differentiate these strains from complex communities and confirm their presence after several weeks in natural environments. We provide two examples of applications using the pigments to trace strains following introduction into plant tissues or to produce a reporter strain for extracellular nitrogen compound sensing. We recognize that this tool could have far broader utility in other applications and microbes, and describe the methodology for use by the greater scientific community.</p>","PeriodicalId":18573,"journal":{"name":"MicrobiologyOpen","volume":"13 4","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mbo3.1425","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141580220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
G protein-coupled estrogen receptor agonist G-1 decreases ADAM10 levels and NLRP3-inflammasome component activation in response to Staphylococcus aureus alpha-hemolysin G蛋白偶联雌激素受体激动剂G-1可降低金黄色葡萄球菌α-溶血素作用下的ADAM10水平和NLRP3-炎症小体成分活化。
IF 3.4 3区 生物学
MicrobiologyOpen Pub Date : 2024-06-12 DOI: 10.1002/mbo3.1423
Huayu Zheng, Kathleen D. Triplett, Eric R. Prossnitz, Pamela R. Hall, Seth M. Daly
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