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Journal articles on the topic 'Transposon mutant library screening'

Author: Grafiati

Published: 3 June 2025

Last updated: 31 July 2025

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1

Bourgeois, Jacob, and Andrew Camilli. "High-Throughput Mutant Screening inVibrio choleraevia Transposon Sequencing." Cold Spring Harbor Protocols 2023, no. 10 (2023): pdb.prot108185. http://dx.doi.org/10.1101/pdb.prot108185.

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Transposon mutagenesis greatly facilitates the study of gene function in microorganisms ranging from viruses to fungi. Traditionally, one would study individual transposon mutants with interesting phenotypes one mutant at a time. Here, we describe methods for the study of tens of thousands of transposon mutants in parallel in the bacterial pathogenVibrio choleraeusing transposon-sequencing. The first section outlines methods for making a saturated transposon mutant library. The second section outlines methods for massively parallel sequencing of the transposon junctions. The third section outl

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Bourgeois, Jacob, and Andrew Camilli. "High-Throughput Mutant Screening via Transposon Sequencing." Cold Spring Harbor Protocols 2023, no. 10 (2023): pdb.top107867. http://dx.doi.org/10.1101/pdb.top107867.

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Transposon mutagenesis has been the method of choice for genetic screens and selections in bacteria by virtue of the transposon being linked to the disrupted gene, simplifying its identification. Transposon sequencing (Tn-seq) is a high-throughput version of transposon mutant screening, in which massively parallel sequencing is used to simultaneously follow the fitness of all mutants in a complex library. In a single experiment, one can use Tn-seq to interrogate the contribution of all genes of a bacterium to fitness under a condition of interest. Here, we introduce a method to construct a sat

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Hudson, P., T. S. Gorton, L. Papazisi, K. Cecchini, S. Frasca, and S. J. Geary. "Identification of a Virulence-Associated Determinant, Dihydrolipoamide Dehydrogenase (lpd), in Mycoplasma gallisepticum through In Vivo Screening of Transposon Mutants." Infection and Immunity 74, no. 2 (2006): 931–39. http://dx.doi.org/10.1128/iai.74.2.931-939.2006.

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ABSTRACT To effectively analyze Mycoplasma gallisepticum for virulence-associated determinants, the ability to create stable genetic mutations is essential. Global M. gallisepticum mutagenesis is currently limited to the use of transposons. Using the gram-positive transposon Tn4001mod, a mutant library of 110 transformants was constructed and all insertion sites were mapped. To identify transposon insertion points, a unique primer directed outward from the end of Tn4001mod was used to sequence flanking genomic regions. By comparing sequences obtained in this manner to the annotated M. gallisep

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Held, Kiara, Elizabeth Ramage, Michael Jacobs, Larry Gallagher, and Colin Manoil. "Sequence-Verified Two-Allele Transposon Mutant Library for Pseudomonas aeruginosa PAO1." Journal of Bacteriology 194, no. 23 (2012): 6387–89. http://dx.doi.org/10.1128/jb.01479-12.

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ABSTRACTMutant hunts using comprehensive sequence-defined libraries make it possible to identify virtually all of the nonessential functions required for different bacterial processes. However, the success of such screening depends on the accuracy of mutant identification in the mutant library used. To provide a high-quality library forPseudomonas aeruginosaPAO1, we created a sequence-verified collection of 9,437 transposon mutants that provides genome coverage and includes two mutants for most genes. Mutants were cherry-picked from a larger library, colony-purified, and resequenced both indiv

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Chen, Qiang, Hui Wu, and Paula M. Fives-Taylor. "Construction of a Novel Transposon Mutagenesis System Useful in the Isolation of Streptococcus parasanguis Mutants Defective in Fap1 Glycosylation." Infection and Immunity 70, no. 12 (2002): 6534–40. http://dx.doi.org/10.1128/iai.70.12.6534-6540.2002.

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ABSTRACT Streptococcus parasanguis, a primary colonizer of the tooth surface, has long, peritrichous fimbriae. A fimbria-associated protein, Fap1, is identified as an adhesin of S. parasanguis FW213. The mature Fap1 protein is glycosylated, and the glycosylation is required for fimbria biogenesis and bacterial adhesion. Little is known about the mechanism of Fap1 glycosylation due to the lack of identifiable mutants. A novel transposon mutagenesis system was established and used to generate a mutant library. Screening of the library with a monoclonal antibody specific for a glycan epitope of F

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Wiegand, Irith, Alexandra K. Marr, Elena B. M. Breidenstein, Kristen N. Schurek, Patrick Taylor, and Robert E. W. Hancock. "Mutator Genes Giving Rise to Decreased Antibiotic Susceptibility in Pseudomonas aeruginosa." Antimicrobial Agents and Chemotherapy 52, no. 10 (2008): 3810–13. http://dx.doi.org/10.1128/aac.00233-08.

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ABSTRACT Screening of the PA14 genomic transposon mutant library for resistance to ceftazidime, tobramycin, and ciprofloxacin led to the discovery of several mutants that appeared in more than one screen. Testing of the frequency of mutation to ciprofloxacin resistance revealed previously known mutator genes, including mutS and mutL, as well as mutators that have not yet been described for P. aeruginosa, including PA3958 and RadA (PA4609).

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Ho, Peiying, Linh Chi Dam, Wei Ren Ryanna Koh, et al. "Screening of the PA14NR Transposon Mutant Library Identifies Genes Involved in Resistance to Bacteriophage Infection in Pseudomomas aeruginosa." International Journal of Molecular Sciences 25, no. 13 (2024): 7009. http://dx.doi.org/10.3390/ijms25137009.

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Multidrug-resistant P. aeruginosa infections pose a serious public health threat due to the rise in antimicrobial resistance. Phage therapy has emerged as a promising alternative. However, P. aeruginosa has evolved various mechanisms to thwart phage attacks, making it crucial to decipher these resistance mechanisms to develop effective therapeutic strategies. In this study, we conducted a forward-genetic screen of the P. aeruginosa PA14 non-redundant transposon library (PA14NR) to identify dominant-negative mutants displaying phage-resistant phenotypes. Our screening process revealed 78 mutant

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Ghatage, Abhay, Rachna Pandey, and G.R Pathade. "Transposon Mutagenesis for enhancement of nitrogen fixing ability of Rhizobium japonicum infecting Vigna radiata." Ecology, Environment and Conservation 29 (2023): 153–56. http://dx.doi.org/10.53550/eec.2023.v29i03s.030.

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This study developed mutant library of 800 mutants with transposition frequency of 3.7 x 10- 6. Transposon mutagenesis is an easy and highly effective method for generating bacteria with improved characteristics and gene knockouts. The slow growing Rhizobium japonicum isolated from root nodules of locally grown mung bean plant. The isolated strain was subjected for transposon mutagenesis to produce 800 mutants. These 800 mutants were screened for nodulation. After screening of these 800 mutants, the100 mutants showed pink coloured nodulation to mung bean plant. Out of 100 mutants 10 were promi

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Karsi, Attila, Nagihan Gülsoy, Erin Corb, Pradeep R. Dumpala, and Mark L. Lawrence. "High-Throughput Bioluminescence-Based Mutant Screening Strategy for Identification of Bacterial Virulence Genes." Applied and Environmental Microbiology 75, no. 7 (2009): 2166–75. http://dx.doi.org/10.1128/aem.02449-08.

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ABSTRACT A high-throughput bioluminescence screening procedure for identification of virulence genes in bacteria was developed and applied to the fish pathogen Edwardsiella ictaluri. A random transposon mutant library expressing bioluminescence was constructed and robotically arrayed on 384-well plates. Mutants were cultivated and mixed with catfish serum and neutrophils in 96-well plates, and bioluminescence was used to detect mutants that are more susceptible to killing by these host factors. The virulence and vaccine efficacy of selected mutants were determined in channel catfish. Transposo

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Maroncle, Nathalie, Damien Balestrino, Chantal Rich, and Christiane Forestier. "Identification of Klebsiella pneumoniae Genes Involved in Intestinal Colonization and Adhesion Using Signature-Tagged Mutagenesis." Infection and Immunity 70, no. 8 (2002): 4729–34. http://dx.doi.org/10.1128/iai.70.8.4729-4734.2002.

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ABSTRACT Klebsiella pneumoniae is an opportunistic pathogen responsible for nosocomial infections that initially colonize the intestinal tract of patients. Signature-tagged mutagenesis was used to identify genes required for this function. A library of 2,200 mutants was analyzed for the inability of the mutants to survive in a murine model of intestinal colonization and to adhere to human intestinal cells (Int-407) in vitro. Twenty-nine attenuated mutants were selected for further analyses after competition assays against the wild-type strain. Whatever the screening model, most of the transpos

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Throne-Holst, Mimmi, Alexander Wentzel, Trond E. Ellingsen, Hans-Kristian Kotlar, and Sergey B. Zotchev. "Identification of Novel Genes Involved in Long-Chain n-Alkane Degradation by Acinetobacter sp. Strain DSM 17874." Applied and Environmental Microbiology 73, no. 10 (2007): 3327–32. http://dx.doi.org/10.1128/aem.00064-07.

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ABSTRACT Acinetobacter sp. strain DSM 17874 is capable of utilizing n-alkanes with chain lengths ranging from that of decane (C10H22) to that of tetracontane (C40H82) as a sole carbon source. Two genes encoding AlkB-type alkane hydroxylase homologues, designated alkMa and alkMb, have been shown to be involved in the degradation of n-alkanes with chain lengths of from 10 to 20 C atoms in this strain. Here, we describe a novel high-throughput screening method and the screening of a transposon mutant library to identify genes involved in the degradation of n-alkanes with C chain lengths longer th

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Yu, Jie, Yaqi Wang, Dongmei Han, et al. "Identification of Streptococcus mutans genes involved in fluoride resistance by screening of a transposon mutant library." Molecular Oral Microbiology 35, no. 6 (2020): 260–70. http://dx.doi.org/10.1111/omi.12316.

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Pahlevi, Muhammad Reza, Keiji Murakami, Yuka Hiroshima, Akikazu Murakami, and Hideki Fujii. "pruR and PA0065 Genes Are Responsible for Decreasing Antibiotic Tolerance by Autoinducer Analog-1 (AIA-1) in Pseudomonas aeruginosa." Antibiotics 11, no. 6 (2022): 773. http://dx.doi.org/10.3390/antibiotics11060773.

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Pseudomonas aeruginosa infection is considered a high-risk nosocomial infection and is very difficult to eradicate because of its tolerance to antibiotic treatment. A new compound, autoinducer analog-1 (AIA-1), has been demonstrated to reduce antibiotic tolerance, but its mechanisms remain unknown. This study aimed to investigate the mechanisms of AIA-1 in the antibiotic tolerance of P. aeruginosa. A transposon mutant library was constructed using miniTn5pro, and screening was performed to isolate high tolerant mutants upon exposure to biapenem and AIA-1. We constructed a deletion mutant and c

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Lanckriet, A., L. Timbermont, L. J. Happonen, et al. "Generation of Single-Copy Transposon Insertions in Clostridium perfringens by Electroporation of Phage Mu DNA Transposition Complexes." Applied and Environmental Microbiology 75, no. 9 (2009): 2638–42. http://dx.doi.org/10.1128/aem.02214-08.

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ABSTRACT Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was s

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Ewann, Fanny, Mary Jackson, Kevin Pethe, et al. "Transient Requirement of the PrrA-PrrB Two-Component System for Early Intracellular Multiplication of Mycobacterium tuberculosis." Infection and Immunity 70, no. 5 (2002): 2256–63. http://dx.doi.org/10.1128/iai.70.5.2256-2263.2002.

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ABSTRACT Adaptive regulation of gene expression in response to environmental changes is a general property of bacterial pathogens. By screening an ordered transposon mutagenesis library of Mycobacterium tuberculosis, we have identified three mutants containing a transposon in the coding sequence or in the 5′ regions of genes coding for two-component signal transduction systems (trcS, regX3, prrA). The intracellular multiplication capacity of the three mutants was investigated in mouse bone marrow-derived macrophages. Only the prrA mutant showed a defect in intracellular growth during the early

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Cartman, Stephen T., and Nigel P. Minton. "A mariner-Based Transposon System for In Vivo Random Mutagenesis of Clostridium difficile." Applied and Environmental Microbiology 76, no. 4 (2009): 1103–9. http://dx.doi.org/10.1128/aem.02525-09.

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ABSTRACT Understanding the molecular basis of Clostridium difficile infection is a prerequisite to the development of effective countermeasures. Although there are methods for constructing gene-specific mutants of C. difficile, currently there is no effective method for generating libraries of random mutants. In this study, we developed a novel mariner-based transposon system for in vivo random mutagenesis of C. difficile R20291, the BI/NAP1/027 epidemic strain at the center of the C. difficile outbreaks in Stoke Mandeville, United Kingdom, in 2003 to 2004 and 2004 to 2005. Transposition occur

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Overhage, Joerg, Shawn Lewenza, Alexandra K. Marr, and Robert E. W. Hancock. "Identification of Genes Involved in Swarming Motility Using a Pseudomonas aeruginosa PAO1 Mini-Tn5-lux Mutant Library." Journal of Bacteriology 189, no. 5 (2006): 2164–69. http://dx.doi.org/10.1128/jb.01623-06.

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ABSTRACT During a screening of a mini-Tn5-luxCDABE transposon mutant library of Pseudomonas aeruginosa PAO1 for alterations in swarming motility, 36 mutants were identified with Tn5 insertions in genes for the synthesis or function of flagellin and type IV pilus, in genes for the Xcp-related type II secretion system, and in regulatory, metabolic, chemosensory, and hypothetical genes with unknown functions. These mutants were differentially affected in swimming and twitching motility but in most cases had only a minor additional motility defect. Our data provide evidence that swarming is a more

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Alexander, David C., Joses R. W. Jones, and Jun Liu. "A Rifampin-Hypersensitive Mutant Reveals Differences between Strains of Mycobacterium smegmatis and Presence of a Novel Transposon, IS1623." Antimicrobial Agents and Chemotherapy 47, no. 10 (2003): 3208–13. http://dx.doi.org/10.1128/aac.47.10.3208-3213.2003.

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ABSTRACT Rifampin is a front-line antibiotic for the treatment of tuberculosis. Infections caused by rifampin- and multidrug-resistant Mycobacterium tuberculosis strains are difficult to treat and contribute to a poor clinical outcome. Rifampin resistance most often results from mutations in rpoB. However, some drug-resistant strains have rpoB alleles that encode the phenotype for susceptibility. Similarly, non-M. tuberculosis mycobacteria exhibit higher levels of baseline resistance to rifampin, despite the presence of rpoB alleles that encode the phenotype for susceptibility. To identify oth

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Palanisamy, Rajesh, Yan Zhang, and Guoquan Zhang. "Role of Type 4B Secretion System Protein, IcmE, in the Pathogenesis of Coxiella burnetii." Pathogens 13, no. 5 (2024): 405. http://dx.doi.org/10.3390/pathogens13050405.

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Coxiella burnetii is an obligate intracellular Gram-negative bacterium that causes Q fever, a life-threatening zoonotic disease. C. burnetii replicates within an acidified parasitophorous vacuole derived from the host lysosome. The ability of C. burnetii to replicate and achieve successful intracellular life in the cell cytosol is vastly dependent on the Dot/Icm type 4B secretion system (T4SSB). Although several T4SSB effector proteins have been shown to be important for C. burnetii virulence and intracellular replication, the role of the icmE protein in the host–C. burnetii interaction has no

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Pětrošová, Helena, and Mathieu Picardeau. "Screening of a Leptospira biflexa Mutant Library To Identify Genes Involved in Ethidium Bromide Tolerance." Applied and Environmental Microbiology 80, no. 19 (2014): 6091–103. http://dx.doi.org/10.1128/aem.01619-14.

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ABSTRACTLeptospiraspp. are spirochete bacteria comprising both pathogenic and free-living species. The saprophyteL. biflexais a model bacterium for studying leptospiral biology due to relative ease of culturing and genetic manipulation. In this study, we constructed a library of 4,996 random transposon mutants inL. biflexa. We screened the library for increased susceptibility to the DNA intercalating agent, ethidium bromide (EtBr), in order to identify genetic determinants that reduceL. biflexasusceptibility to antimicrobial agents. By phenotypic screening, using subinhibitory EtBr concentrati

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Shin, Sung Jae, Chia-wei Wu, Howard Steinberg, and Adel M. Talaat. "Identification of Novel Virulence Determinants in Mycobacterium paratuberculosis by Screening a Library of Insertional Mutants." Infection and Immunity 74, no. 7 (2006): 3825–33. http://dx.doi.org/10.1128/iai.01742-05.

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ABSTRACT Johne's disease, caused by Mycobacterium paratuberculosis infection, is a worldwide problem for the dairy industry and has a possible involvement in Crohn's disease in humans. To identify virulence determinants of this economically important pathogen, a library of 5,060 transposon mutants was constructed using Tn5367 insertion mutagenesis, followed by large-scale sequencing to identify disrupted genes. In this report, 1,150 mutants were analyzed and 970 unique insertion sites were identified. Sequence analysis of the disrupted genes indicated that the insertion of Tn5367 was more prev

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Rhodes, Katherine A., Man Cheong Ma, María A. Rendón, and Magdalene So. "Neisseria genes required for persistence identified via in vivo screening of a transposon mutant library." PLOS Pathogens 18, no. 5 (2022): e1010497. http://dx.doi.org/10.1371/journal.ppat.1010497.

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The mechanisms used by human adapted commensal Neisseria to shape and maintain a niche in their host are poorly defined. These organisms are common members of the mucosal microbiota and share many putative host interaction factors with Neisseria meningitidis and Neisseria gonorrhoeae. Evaluating the role of these shared factors during host carriage may provide insight into bacterial mechanisms driving both commensalism and asymptomatic infection across the genus. We identified host interaction factors required for niche development and maintenance through in vivo screening of a transposon muta

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Fodor, Barna, Gábor Rákhely, Ákos T. Kovács та Kornél L. Kovács. "Transposon Mutagenesis in Purple Sulfur Photosynthetic Bacteria: Identification of hypF, Encoding a Protein Capable of Processing [NiFe] Hydrogenases in α, β, and γ Subdivisions of the Proteobacteria". Applied and Environmental Microbiology 67, № 6 (2001): 2476–83. http://dx.doi.org/10.1128/aem.67.6.2476-2483.2001.

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ABSTRACT A random transposon-based mutagenesis system was optimized for the purple sulfur phototrophic bacterium Thiocapsa roseopersicina BBS. Screening for hydrogenase-deficient phenotypes resulted in the isolation of six independent mutants in a mini-Tn5 library. One of the mutations was in a gene showing high amino acid sequence similarity to HypF proteins in other organisms. Inactivation of hydrogen uptake activity in thehypF-deficient mutant resulted in a dramatic increase in the hydrogen evolution capacity of T. roseopersicinaunder nitrogen-fixing conditions. This mutant is therefore a p

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Zhang, Xinglin, Maat Vincent de, Prieto Ana M. Guzmán, et al. "RNA-seq and Tn-seq reveal fitness determinants of vancomycin-resistant Enterococcus faecium during growth in human serum." BMC Genomics 18, no. 1 (2017): 893. https://doi.org/10.1186/s12864-017-4299-9.

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<strong>Background: </strong>The Gram-positive bacterium <i>Enterococcus faecium</i> is a commensal of the human gastrointestinal tract and a frequent cause of bloodstream infections in hospitalized patients. The mechanisms by which <i>E. faecium</i> can survive and grow in blood during an infection have not yet been characterized. Here, we identify genes that contribute to growth of <i>E. faecium</i> in human serum through transcriptome profiling (RNA-seq) and a high-throughput transposon mutant library sequencing approach (Tn-seq).<strong>Results: </strong>We first sequenced the genome of <i

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Li, Xiangzhen, Qingwei Luo, Neil Q. Wofford, et al. "A Molybdopterin Oxidoreductase Is Involved in H2 Oxidation in Desulfovibrio desulfuricans G20." Journal of Bacteriology 191, no. 8 (2009): 2675–82. http://dx.doi.org/10.1128/jb.01814-08.

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ABSTRACT Three mutants deficient in hydrogen/formate uptake were obtained through screening of a transposon mutant library containing 5,760 mutants of Desulfovibrio desulfuricans G20. Mutations were in the genes encoding the type I tetraheme cytochrome c 3 (cycA), Fe hydrogenase (hydB), and molybdopterin oxidoreductase (mopB). Mutations did not decrease the ability of cells to produce H2 or formate during growth. Complementation of the cycA and mopB mutants with a plasmid carrying the intact cycA and/or mopB gene and the putative promoter from the parental strain allowed the recovery of H2 upt

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Fan, Jingyan, Lelin Zhao, Qiao Hu, et al. "Screening for Virulence-Related Genes via a Transposon Mutant Library of Streptococcus suis Serotype 2 Using a Galleria mellonella Larvae Infection Model." Microorganisms 10, no. 5 (2022): 868. http://dx.doi.org/10.3390/microorganisms10050868.

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Streptococcus suis (S. suis) is a zoonotic bacterial pathogen causing lethal infections in pigs and humans. Identification of virulence-related genes (VRGs) is of great importance in understanding the pathobiology of a bacterial pathogen. To identify novel VRGs, a transposon (Tn) mutant library of S. suis strain SC19 was constructed in this study. The insertion sites of approximately 1700 mutants were identified by Tn-seq, which involved 417 different genes. A total of 32 attenuated strains were identified from the library by using a Galleria mellonella larvae infection model, and 30 novel VRG

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Pei, Xiaomeng, Mingxing Liu, Hong Zhou, and Hongjie Fan. "Screening for phagocytosis resistance-related genes via a transposon mutant library of Streptococcus suis serotype 2." Virulence 11, no. 1 (2020): 825–38. http://dx.doi.org/10.1080/21505594.2020.1782088.

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Tareen, A. Malik, Javid Iqbal Dasti, Andreas E. Zautner, Uwe Groß, and Raimond Lugert. "Sulphite : cytochrome c oxidoreductase deficiency in Campylobacter jejuni reduces motility, host cell adherence and invasion." Microbiology 157, no. 6 (2011): 1776–85. http://dx.doi.org/10.1099/mic.0.045567-0.

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Campylobacter jejuni lacks the enzyme phosphofructokinase and, consequently, is incapable of metabolizing glucose. Instead, the pathogen uses a number of other chemicals to serve as electron donors. Like chemolithotrophic bacteria, C. jejuni is able to respire sulphite in the presence of a sulphite : cytochrome c oxidoreductase (SOR) that is encoded by the genes cj0004c and cj0005c; the former encodes a monohaem cytochrome c oxidoreductase and the latter a molybdopterin oxidoreductase. After screening of a transposon-based mutant library, we identified a mutant with an insertion in gene cj0005

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Lindemann, Stephen R., Kaitian Peng, Matthew E. Long, et al. "Francisella tularensisSchu S4 O-Antigen and Capsule Biosynthesis Gene Mutants Induce Early Cell Death in Human Macrophages." Infection and Immunity 79, no. 2 (2010): 581–94. http://dx.doi.org/10.1128/iai.00863-10.

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ABSTRACTFrancisella tularensisis capable of rampant intracellular growth and causes a potentially fatal disease in humans. Whereas many mutational studies have been performed with avirulent strains ofFrancisella, relatively little has been done with strains that cause human disease. We generated a near-saturating transposon library in the virulent strain Schu S4, which was subjected to high-throughput screening by transposon site hybridization through primary human macrophages, negatively selecting 202 genes. Of special note were genes in a locus of theFrancisellachromosome,FTT1236,FTT1237, an

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Yamazaki, Yoshitaka, Lia Danelishvili, Martin Wu, Molly MacNab, and Luiz E. Bermudez. "Mycobacterium avium Genes Associated with the Ability To Form a Biofilm." Applied and Environmental Microbiology 72, no. 1 (2006): 819–25. http://dx.doi.org/10.1128/aem.72.1.819-825.2006.

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ABSTRACT Mycobacterium avium is widely distributed in the environment, and it is chiefly found in water and soil. M. avium, as well as Mycobacterium smegmatis, has been recognized to produce a biofilm or biofilm-like structure. We screened an M. avium green fluorescent protein (GFP) promoter library in M. smegmatis for genes involved in biofilm formation on polyvinyl chloride (PVC) plates. Clones associated with increased GFP expression ≥2.0-fold over the baseline were sequenced. Seventeen genes, most encoding proteins of the tricarboxylic acid (TCA) cycle and GDP-mannose and fatty acid biosyn

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Coutinho, Bruna G., Danilo Licastro, Lucia Mendonça-Previato, Miguel Cámara, and Vittorio Venturi. "Plant-Influenced Gene Expression in the Rice Endophyte Burkholderia kururiensis M130." Molecular Plant-Microbe Interactions® 28, no. 1 (2015): 10–21. http://dx.doi.org/10.1094/mpmi-07-14-0225-r.

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Burkholderia kururiensis M130 is one of the few rice endophytic diazotrophic bacteria identified thus far which is able to enhance growth of rice. To date, very little is known of how strain M130 and other endophytes enter and colonize plants. Here, we identified genes of strain M130 that are differentially regulated in the presence of rice plant extract. A genetic screening of a promoter probe transposon mutant genome bank and RNAseq analysis were performed. The screening of 10,100 insertions of the genomic transposon reporter library resulted in the isolation of 61 insertions displaying diff

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Legerme, Georgio, Evan Yang, Rianne Esquivel, Saija Kiljunen, Harri Savilahti, and Mechthild Pohlschroder. "Screening of a Haloferax volcanii Transposon Library Reveals Novel Motility and Adhesion Mutants." Life 6, no. 4 (2016): 41. http://dx.doi.org/10.3390/life6040041.

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Xiao, Xia, Yi Li, Liang Li, and Yan Xiong. "Identification of Methicillin-Resistant Staphylococcus aureus (MRSA) Genetic Factors Involved in Human Endothelial Cells Damage, an Important Phenotype Correlated with Persistent Endovascular Infection." Antibiotics 11, no. 3 (2022): 316. http://dx.doi.org/10.3390/antibiotics11030316.

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Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of life-threatening endovascular infections. Endothelial cell (EC) damage is a key factor in the pathogenesis of these syndromes. However, genetic factors related to the EC damage have not been well studied. This study aims to identify genetic determinants that impact human EC damage by screening the genome-wide Nebraska Transposon Mutant Library (NTML). A well-established MTT assay was used to test the in vitro damage of human EC cell line (HMEC-1) caused by each mutant strain in the NTML. We first confirmed some global reg

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Liu, Yue, Xue Xia, and Fengyi Wan. "Random mutagenesis unveils novel host-pathogen interactions during colonic bacterial infections in immunocompromised hosts." Journal of Immunology 200, no. 1_Supplement (2018): 117.33. http://dx.doi.org/10.4049/jimmunol.200.supp.117.33.

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Abstract Attaching/Effacing (A/E) pathogens including enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC), and the rodent equivalent Citrobacter rodentium (CR) are important causative agents of foodborne diseases. A/E pathogen infections cause severe morbidity and mortality in immunocompromised hosts with low interleukin-22 (IL-22); however, the crucial host-pathogen interactions and the pivotal A/E virulence proteins (effectors) under immunocompromised conditions, remain elusive. We conducted a functional screening of a CR mutant library consisting of ~2,000 mutant stra

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Chee, Jessica. "Identifying genes associated with biofilm production in Pseudomonas aeruginosa." Meducator 1, no. 33 (2018): 19–22. http://dx.doi.org/10.15173/m.v1i33.1796.

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Pseudomonas aeruginosa is an opportunistic pathogen associated with a variety of life-threatening diseases. Infections caused by P. aeruginosa can be nearly untreatable because of its multidrug-resistance. One of the characteristics of P. aeruginosa that helps it survive in high drug concentrations is its ability to form biofilms–large communities of cells encompassed by extracellular polymeric substances that defend against many antibiotics. In fact, sub-minimum inhibitory concentrations of antibiotics stimulate biofilm production. This project aims to identify genes associated with biofilm i

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Hingston, Patricia A., Marta J. Piercey, and Lisbeth Truelstrup Hansen. "Genes Associated with Desiccation and Osmotic Stress in Listeria monocytogenes as Revealed by Insertional Mutagenesis." Applied and Environmental Microbiology 81, no. 16 (2015): 5350–62. http://dx.doi.org/10.1128/aem.01134-15.

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ABSTRACTListeria monocytogenesis a foodborne pathogen whose survival in food processing environments may be associated with its tolerance to desiccation. To probe the molecular mechanisms used by this bacterium to adapt to desiccation stress, a transposon library of 11,700L. monocytogenesmutants was screened, using a microplate assay, for strains displaying increased or decreased desiccation survival (43% relative humidity, 15°C) in tryptic soy broth (TSB). The desiccation phenotypes of selected mutants were subsequently assessed on food-grade stainless steel (SS) coupons in TSB plus 1% glucos

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Hobom, Urs, Wolfram Brune, Martin Messerle, Gabriele Hahn, and Ulrich H. Koszinowski. "Fast Screening Procedures for Random Transposon Libraries of Cloned Herpesvirus Genomes: Mutational Analysis of Human Cytomegalovirus Envelope Glycoprotein Genes." Journal of Virology 74, no. 17 (2000): 7720–29. http://dx.doi.org/10.1128/jvi.74.17.7720-7729.2000.

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ABSTRACT We have cloned the human cytomegalovirus (HCMV) genome as an infectious bacterial artificial chromosome (BAC) in Escherichia coli. Here, we have subjected the HCMV BAC to random transposon (Tn) mutagenesis using a Tn1721-derived insertion sequence and have provided the conditions for excision of the BAC cassette. We report on a fast and efficient screening procedure for a Tn insertion library. Bacterial clones containing randomly mutated full-length HCMV genomes were transferred into 96-well microtiter plates. A PCR screening method based on two Tn primers and one primer specific for

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Wu, Weihui, Hassan Badrane, Shiwani Arora, Henry V. Baker, and Shouguang Jin. "MucA-Mediated Coordination of Type III Secretion and Alginate Synthesis in Pseudomonas aeruginosa." Journal of Bacteriology 186, no. 22 (2004): 7575–85. http://dx.doi.org/10.1128/jb.186.22.7575-7585.2004.

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ABSTRACT The type III secretion system (T3SS) of Pseudomonas aeruginosa is an important virulence factor. The T3SS of P. aeruginosa can be induced by a low calcium signal or upon direct contact with the host cells. The exact pathway of signal sensing and T3SS activation is not clear. By screening a transposon insertion mutant library of the PAK strain, mutation in the mucA gene was found to cause repression of T3SS expression under both type III-inducing and -noninducing conditions. Mutation in the mucA gene is known to cause alginate overproduction, resulting in a mucoid phenotype. Alginate p

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Geng, Haoyu, Linyan Luo, Jian Zhang, Jingying Gao, Shizhong Geng, and Paul Barrow. "Identification of a Novel Regulatory Gene, trmE, that Orchestrates Salmonella Flagellar Synthesis and Virulence." Microorganisms 13, no. 7 (2025): 1455. https://doi.org/10.3390/microorganisms13071455.

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It is well established that flagella play a critical role in bacterial motility and virulence, and the genes associated with flagellar synthesis and regulation have been extensively characterized. In this study, we identified the trmE gene as a novel modulator of flagellar synthesis in Salmonella Enteritidis. A transposon (Tn5) mutant library of Salmonella Enteritidis (SE) was constructed through bacterial conjugation, followed by screening for motility-deficient mutants. Among 1321 mutants screened, C50041trmE::Tn5 exhibited reduced motility. To validate this phenotype, we constructed C50041Δ

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Fernández, Lucía, Carolina Álvarez-Ortega, Irith Wiegand, et al. "Characterization of the Polymyxin B Resistome of Pseudomonas aeruginosa." Antimicrobial Agents and Chemotherapy 57, no. 1 (2012): 110–19. http://dx.doi.org/10.1128/aac.01583-12.

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ABSTRACTMultidrug resistance inPseudomonas aeruginosais increasingly becoming a threat for human health. Indeed, some strains are resistant to almost all currently available antibiotics, leaving very limited choices for antimicrobial therapy. In many such cases, polymyxins are the only available option, although as their utilization increases so does the isolation of resistant strains. In this study, we screened a comprehensive PA14 mutant library to identify genes involved in changes of susceptibility to polymyxin B inP. aeruginosa. Surprisingly, our screening revealed that the polymyxin B re

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Chapman, Christine, and Louis S. Tisa. "Identification and characterization of Photorhabdus temperata mutants altered in hemolysis and virulence." Canadian Journal of Microbiology 62, no. 8 (2016): 657–67. http://dx.doi.org/10.1139/cjm-2016-0102.

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Photorhabdus temperata is a symbiont of the entomopathogenic nematode Heterorhabditis bacteriophora and an insect pathogen. This bacterium produces a wide variety of virulence factors and hemolytic activity. The goal of this study was to identify hemolysin-defective mutants and test their virulence. A genetic approach was used to identify mutants with altered hemolytic activity by screening a library of 10 000 P. temperata transposon mutants. Three classes of mutants were identified: (i) defective (no hemolytic activity), (ii) delayed (delayed initiation of hemolytic activity), and (iii) early

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Sabbagh, Sébastien C., Christine Lepage, Michael McClelland, and France Daigle. "Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library." PLoS ONE 7, no. 5 (2012): e36643. http://dx.doi.org/10.1371/journal.pone.0036643.

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Yamaguchi, Mikiyo, Keiko Sato, Hideharu Yukitake, Yuichiro Noiri, Shigeyuki Ebisu, and Koji Nakayama. "A Porphyromonas gingivalis Mutant Defective in a Putative Glycosyltransferase Exhibits Defective Biosynthesis of the Polysaccharide Portions of Lipopolysaccharide, Decreased Gingipain Activities, Strong Autoaggregation, and Increased Biofilm Formation." Infection and Immunity 78, no. 9 (2010): 3801–12. http://dx.doi.org/10.1128/iai.00071-10.

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ABSTRACT The Gram-negative anaerobic bacterium Porphyromonas gingivalis is a major pathogen in periodontal disease, one of the biofilm-caused infectious diseases. The bacterium possesses potential virulence factors, including fimbriae, proteinases, hemagglutinin, lipopolysaccharide (LPS), and outer membrane vesicles, and some of these factors are associated with biofilm formation; however, the precise mechanism of biofilm formation is still unknown. Colonial pigmentation of the bacterium on blood agar plates is related to its virulence. In this study, we isolated a nonpigmented mutant that had

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Hood, M. Indriati, Kyle W. Becker, Christelle M. Roux, Paul M. Dunman, and Eric P. Skaar. "Genetic Determinants of Intrinsic Colistin Tolerance in Acinetobacter baumannii." Infection and Immunity 81, no. 2 (2012): 542–51. http://dx.doi.org/10.1128/iai.00704-12.

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ABSTRACTAcinetobacter baumanniiis a leading cause of multidrug-resistant infections worldwide. This organism poses a particular challenge due to its ability to acquire resistance to new antibiotics through adaptation or mutation. This study was undertaken to determine the mechanisms governing the adaptability ofA. baumanniito the antibiotic colistin. Screening of a transposon mutant library identified over 30 genes involved in inducible colistin resistance inA. baumannii. One of the genes identified waslpsB, which encodes a glycosyltransferase involved in lipopolysaccharide (LPS) synthesis. We

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Hegde, Shivanand, Shrilakshmi Hegde, Martina Zimmermann, et al. "Simultaneous Identification of Potential Pathogenicity Factors of Mycoplasma agalactiae in the Natural Ovine Host by Negative Selection." Infection and Immunity 83, no. 7 (2015): 2751–61. http://dx.doi.org/10.1128/iai.00403-15.

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Mycoplasmas possess complex pathogenicity determinants that are largely unknown at the molecular level.Mycoplasma agalactiaeserves as a useful model to study the molecular basis of mycoplasma pathogenicity. The generation andin vivoscreening of a transposon mutant library ofM. agalactiaewere employed to unravel its host colonization factors. Tn4001mod mutants were sequenced using a novel sequencing method, and functionally heterogeneous pools containing 15 to 19 selected mutants were screened simultaneously through two successive cycles of sheep intramammary infections. A PCR-based negative se

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Schwan, William R., Silvija N. Coulter, Eva Y. W. Ng, et al. "Identification and Characterization of the PutP Proline Permease That Contributes to In Vivo Survival ofStaphylococcus aureus in Animal Models." Infection and Immunity 66, no. 2 (1998): 567–72. http://dx.doi.org/10.1128/iai.66.2.567-572.1998.

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ABSTRACT Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscesses, endocarditis, and other infectious syndromes. A signature-tagged mutagenesis (STM) system was adapted for use in studying the genes required for in vivo survival of S. aureus. An STM library was ultimately created in S. aureus RN6390, with Tn917 being used to create the transposon mutations. Pools of S. aureusRN6390 mutants were screened in mouse abscess, bacteremia, and wound infection models for growth attenuation after in vivo passage. One of the mutants that was identified di

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Chen, You, Yong-Ick Kim, Shannon R. Mackey, C. Kay Holtman, Andy LiWang, and Susan S. Golden. "A Novel Allele of kaiA Shortens the Circadian Period and Strengthens Interaction of Oscillator Components in the Cyanobacterium Synechococcus elongatus PCC 7942." Journal of Bacteriology 191, no. 13 (2009): 4392–400. http://dx.doi.org/10.1128/jb.00334-09.

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ABSTRACT The basic circadian oscillator of the unicellular fresh water cyanobacterium Synechococcus elongatus PCC 7942, the model organism for cyanobacterial circadian clocks, consists of only three protein components: KaiA, KaiB, and KaiC. These proteins, all of which are homomultimers, periodically interact to form large protein complexes with stoichiometries that depend on the phosphorylation state of KaiC. KaiA stimulates KaiC autophosphorylation through direct physical interactions. Screening a library of S. elongatus transposon mutants for circadian clock phenotypes uncovered an atypical

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Peng, Yongchong, Tian Tang, Qianqian Li, et al. "Mycobacterium tuberculosis FadD18 Promotes Proinflammatory Cytokine Secretion to Inhibit the Intracellular Survival of Bacillus Calmette–Guérin." Cells 13, no. 12 (2024): 1019. http://dx.doi.org/10.3390/cells13121019.

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Mycobacterium tuberculosis causes 6.4 million cases of tuberculosis and claims 1.6 million lives annually. Mycobacterial adhesion, invasion of host cells, and subsequent intracellular survival are crucial for the infection and dissemination process, yet the cellular mechanisms underlying these phenomena remain poorly understood. This study created a Bacillus Calmette–Guérin (BCG) transposon library using a MycomarT7 phage carrying a Himar1 Mariner transposon to identify genes related to mycobacteria adhesion and invasion. Using adhesion and invasion model screening, we found that the mutant st

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Sun, Yang, Wenjing Shuai, Lanmengya Nie, Xiangfei Li, and Ling Jiang. "Investigating the Role of OrbF in Biofilm Biosynthesis and Regulation of Biofilm-Associated Genes in Bacillus cereus BC1." Foods 13, no. 5 (2024): 638. http://dx.doi.org/10.3390/foods13050638.

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Bacillus cereus (B. cereus), a prevalent foodborne pathogen, constitutes a substantial risk to food safety due to its pronounced resilience under adverse environmental conditions such as elevated temperatures and ultraviolet radiation. This resilience can be attributed to its capacity for biofilm synthesis and sustained high viability. Our research aimed to elucidate the mechanisms governing biofilm biosynthesis in B. cereus. To this end, we constructed a 5088-mutant library of the B. cereus strain BC1 utilizing the transposon TnYLB-1. Systematic screening of this library yielded mutants exhib

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Spiers, Andrew J., Sophie G. Kahn, John Bohannon, Michael Travisano, and Paul B. Rainey. "Adaptive Divergence in Experimental Populations of Pseudomonas fluorescens. I. Genetic and Phenotypic Bases of Wrinkly Spreader Fitness." Genetics 161, no. 1 (2002): 33–46. http://dx.doi.org/10.1093/genetics/161.1.33.

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Abstract A central feature of all adaptive radiations is morphological divergence, but the phenotypic innovations that are responsible are rarely known. When selected in a spatially structured environment, populations of the bacterium Pseudomonas fluorescens rapidly diverge. Among the divergent morphs is a mutant type termed “wrinkly spreader” (WS) that colonizes a new niche through the formation of self-supporting biofilms. Loci contributing to the primary phenotypic innovation were sought by screening a WS transposon library for niche-defective (WS-) mutants. Detailed analysis of one group o

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