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Transposon mutagenesis

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https://www.readbyqxmd.com/read/29209072/free-sepf-interferes-with-recruitment-of-late-cell-division-proteins
#1
Yongqiang Gao, Michaela Wenzel, Martijs J Jonker, Leendert W Hamoen
The conserved cell division protein SepF aligns polymers of FtsZ, the key cell division protein in bacteria, during synthesis of the (Fts)Z-ring at midcell, the first stage in cytokinesis. In addition, SepF acts as a membrane anchor for the Z-ring. Recently, it was shown that SepF overexpression in Mycobacterium smegmatis blocks cell division. Why this is the case is not known. Surprisingly, we found in Bacillus subtilis that SepF overproduction does not interfere with Z-ring assembly, but instead blocks assembly of late division proteins responsible for septum synthesis...
December 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29208119/comparative-analysis-of-the-burkholderia-cenocepacia-k56-2-essential-genome-reveals-cell-envelope-functions-that-are-uniquely-required-for-survival-in-species-of-the-genus-burkholderia
#2
April S Gislason, Keith Turner, Mike Domaratzki, Silvia T Cardona
Burkholderia cenocepacia K56-2 belongs to the Burkholderia cepacia complex, a group of Gram-negative opportunistic pathogens that have large and dynamic genomes. In this work, we identified the essential genome of B. cenocepacia K56-2 using high-density transposon mutagenesis and insertion site sequencing (Tn-seq circle). We constructed a library of one million transposon mutants and identified the transposon insertions at an average of one insertion per 27 bp. The probability of gene essentiality was determined by comparing of the insertion density per gene with the variance of neutral datasets generated by Monte Carlo simulations...
November 2017: Microbial Genomics
https://www.readbyqxmd.com/read/29205551/regulatory-genes-coordinating-antibiotic-induced-changes-in-promoter-activity-and-early-transcriptional-termination-of-the-mycobacterial-intrinsic-resistance-gene-whib7
#3
Ján Burian, Charles J Thompson
Diseases caused by various Mycobacterium sp., especially Mycobacterium tuberculosis, are a major burden on global health care. Due to high intrinsic antibiotic resistance, treatment options are severely limited. In mycobacteria, WhiB7 coordinates intrinsic resistance to a broad range of antibiotics. While WhiB7 has been established as an auto-regulatory transcriptional activator, the signals and genes needed to induce its expression are poorly understood. Using Mycobacterium smegmatis as a model, we coupled transposon mutagenesis and next generation sequencing with WhiB7-specific antibiotic selection to identify genes that contribute to WhiB7 regulation and function...
December 4, 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/29188707/increasing-extracellular-free-fatty-acid-production-in-escherichia-coli-by-disrupting-membrane-transport-systems
#4
Kwang Soo Shin, Sung Kuk Lee
Transposon mutagenesis was used to identify three mutants of E. coli that exhibited increased free fatty acid (FFA) production, which resulted from the disruption of genes related to membrane transport. Deletion of envR, gusC, and mdlA individually in a recombinant E. coli strain resulted in 1.4-, 1.8-, and 1.2-fold increases in total FFA production, respectively. In particular, deletion of envR increased the percentage of extracellular FFA to 46%, compared with 29% for the control strain. Multiple deletion of envR, gusC, mdlA, ompF, and fadL had a synergistic effect on FFA production, resulting in high extracellular FFA production, comprising up to 50% of total FFA production...
November 30, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29186514/permissive-zones-for-the-centromere-binding-protein-parb-on-the-caulobacter-crescentus-chromosome
#5
Ngat T Tran, Clare E Stevenson, Nicolle F Som, Anyarat Thanapipatsiri, Adam S B Jalal, Tung B K Le
Proper chromosome segregation is essential in all living organisms. In Caulobacter crescentus, the ParA-ParB-parS system is required for proper chromosome segregation and cell viability. The bacterial centromere-like parS DNA locus is the first to be segregated following chromosome replication. parS is bound by ParB protein, which in turn interacts with ParA to partition the ParB-parS nucleoprotein complex to each daughter cell. Here, we investigated the genome-wide distribution of ParB on the Caulobacter chromosome using a combination of in vivo chromatin immunoprecipitation (ChIP-seq) and in vitro DNA affinity purification with deep sequencing (IDAP-seq)...
November 23, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29184019/gut-symbiont-bacteroides-fragilis-secretes-a-eukaryotic-like-ubiquitin-protein-that-mediates-intraspecies-antagonism
#6
Maria Chatzidaki-Livanis, Michael J Coyne, Kevin G Roelofs, Rahul R Gentyala, Jarreth M Caldwell, Laurie E Comstock
Human gut Bacteroides species produce different types of toxins that antagonize closely related members of the gut microbiota. Some are toxic effectors delivered by type VI secretion systems, and others are non-contact-dependent secreted antimicrobial proteins. Many strains of Bacteroides fragilis secrete antimicrobial molecules, but only one of these toxins has been described to date (Bacteroidales secreted antimicrobial protein 1 [BSAP-1]). In this study, we describe a novel secreted protein produced by B...
November 28, 2017: MBio
https://www.readbyqxmd.com/read/29184015/molecular-basis-of-bacterial-longevity
#7
Kieran B Pechter, Liang Yin, Yasuhiro Oda, Larry Gallagher, Jianming Yang, Colin Manoil, Caroline S Harwood
It is well known that many bacteria can survive in a growth-arrested state for long periods of time, on the order of months or even years, without forming dormant structures like spores or cysts. How is such longevity possible? What is the molecular basis of such longevity? Here we used the Gram-negative phototrophic alphaproteobacterium Rhodopseudomonas palustris to identify molecular determinants of bacterial longevity. R. palustris maintained viability for over a month after growth arrest due to nutrient depletion when it was provided with light as a source of energy...
November 28, 2017: MBio
https://www.readbyqxmd.com/read/29164142/pathogenesis-molecular-genetics-and-genomics-of-mycobacterium-avium-subsp-paratuberculosis-the-etiologic-agent-of-johne-s-disease
#8
REVIEW
Govardhan Rathnaiah, Denise K Zinniel, John P Bannantine, Judith R Stabel, Yrjö T Gröhn, Michael T Collins, Raúl G Barletta
Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in ruminants causing chronic diarrhea, malnutrition, and muscular wasting. Neonates and young animals are infected primarily by the fecal-oral route. MAP attaches to, translocates via the intestinal mucosa, and is phagocytosed by macrophages. The ensuing host cellular immune response leads to granulomatous enteritis characterized by a thick and corrugated intestinal wall. We review various tissue culture systems, ileal loops, and mice, goats, and cattle used to study MAP pathogenesis...
2017: Frontiers in Veterinary Science
https://www.readbyqxmd.com/read/29155794/an-array-based-comparative-genomic-hybridization-platform-for-efficient-detection-of-copy-number-variations-in-fast-neutron-induced-medicago-truncatula-mutants
#9
Yuhui Chen, Xianfu Wang, Shunfei Lu, Hongcheng Wang, Shibo Li, Rujin Chen
Mutants are invaluable genetic resources for gene function studies. To generate mutant collections, three types of mutagens can be utilized, including biological such as T-DNA or transposon, chemical such as ethyl methanesulfonate (EMS), or physical such as ionization radiation. The type of mutation observed varies depending on the mutagen used. For ionization radiation induced mutants, mutations include deletion, duplication, or rearrangement. While T-DNA or transposon-based mutagenesis is limited to species that are susceptible to transformation, chemical or physical mutagenesis can be applied to a broad range of species...
November 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29143824/pd-1-is-a-haploinsufficient-suppressor-of-t-cell-lymphomagenesis
#10
Tim Wartewig, Zsuzsanna Kurgyis, Selina Keppler, Konstanze Pechloff, Erik Hameister, Rupert Öllinger, Roman Maresch, Thorsten Buch, Katja Steiger, Christof Winter, Roland Rad, Jürgen Ruland
T cell non-Hodgkin lymphomas are a heterogeneous group of highly aggressive malignancies with poor clinical outcomes. T cell lymphomas originate from peripheral T cells and are frequently characterized by genetic gain-of-function variants in T cell receptor (TCR) signalling molecules. Although these oncogenic alterations are thought to drive TCR pathways to induce chronic proliferation and cell survival programmes, it remains unclear whether T cells contain tumour suppressors that can counteract these events...
November 15, 2017: Nature
https://www.readbyqxmd.com/read/29110195/a-transposition-active-phyllostachys-edulis-long-terminal-repeat-ltr-retrotransposon
#11
Mingbing Zhou, Linlin Liang, Heikki Hänninen
Due to infrequent sexual reproduction, moso bamboo breeding by hybridization is extremely technically difficult. Insertional mutagenesis based on endogenous active transposons may thus serve as an alternative method to create new germplasm of moso bamboo. In the present study, using LTR-STRUC, a full-length intact long terminal repeat (LTR) retrotransposon was identified in the moso bamboo genome and was named PHRE2 (Phyllostachys edulis retrotransposon 2). The 5' and 3' LTR sequences of PHRE2 were highly (98...
November 6, 2017: Journal of Plant Research
https://www.readbyqxmd.com/read/29109185/genome-wide-screens-reveal-new-gene-products-that-influence-genetic-competence-in-streptococcus-mutans
#12
Robert C Shields, Greg O'Brien, Natalie Maricic, Alexandria Kesterson, Megan Grace, Stephen J Hagen, Robert A Burne
A network of genes and at least two peptide signaling molecules tightly control when Streptococcus mutans becomes competent to take up DNA from its environment. Widespread changes in the expression of genes occur when S. mutans is presented with competence signal peptides in vitro, including increased production of the alternative sigma factor, ComX, which activates late competence genes. Still, the way that gene products that are regulated by competence peptides influence DNA uptake and cellular physiology are not well understood...
November 6, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/29101196/genome-wide-identification-of-fitness-factors-in-mastitis-associated-escherichia-coli
#13
Michael A Olson, Timothy W Siebach, Joel S Griffitts, Eric Wilson, David L Erickson
Virulence factors of mammary pathogenic Escherichia coli (MPEC) have not been identified, and it is not known how bacterial gene content influences severity of mastitis. Here, we report the first genome-wide identification of genes that contribute to fitness of MPEC in conditions relevant to the natural history of the disease. A highly virulent clinical isolate (M12) was identified that killed Galleria mellonella at low infectious doses and which replicated to high numbers in mouse mammary glands and spread to spleens...
November 3, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29098331/essential-two-component-systems-regulating-cell-envelope-functions-opportunities-for-novel-antibiotic-therapies
#14
Silvia T Cardona, Matthew Choy, Andrew M Hogan
Novel therapies are urgently needed to alleviate the current crisis of multiple drug-resistant infections. The bacterial signal transduction mechanisms, known as two-component systems (TCSs), are ideal targets of novel inhibitory molecules. Highly restricted to the bacterial world, TCSs control a diverse set of cellular functions, namely virulence, response to cell envelope stress, and drug efflux. Impaired regulation of any of these aspects could affect the susceptibility of bacterial pathogens to antibiotics, which highlights the potential of TCS as targets of antibiotic adjuvant therapies...
November 2, 2017: Journal of Membrane Biology
https://www.readbyqxmd.com/read/29091941/csrb-a-noncoding-regulatory-rna-is-required-for-bara-dependent-expression-of-biocontrol-traits-in-rahnella-aquatilis-hx2
#15
Li Mei, Sanger Xu, Peng Lu, Haiping Lin, Yanbin Guo, Yongjun Wang
BACKGROUND: Rahnella aquatilis is ubiquitous and its certain strains have the applicative potent as a plant growth-promoting rhizobacteria. R. aquatilis HX2 is a biocontrol agent to produce antibacterial substance (ABS) and showed efficient biocontrol against crown gall caused by Agrobacterium vitis on sunflower and grapevine plants. The regulatory network of the ABS production and biocontrol activity is still limited known. METHODOLOGY/PRINCIPAL FINDINGS: In this study, a transposon-mediated mutagenesis strategy was used to investigate the regulators that involved in the biocontrol activity of R...
2017: PloS One
https://www.readbyqxmd.com/read/29091192/modifications-in-the-pmrb-gene-are-the-primary-mechanism-for-the-development-of-chromosomally-encoded-resistance-to-polymyxins-in-uropathogenic-escherichia-coli
#16
Minh-Duy Phan, Nguyen Thi Khanh Nhu, Maud E S Achard, Brian M Forde, Kar Wai Hong, Teik Min Chong, Wai-Fong Yin, Kok-Gan Chan, Nicholas P West, Mark J Walker, David L Paterson, Scott A Beatson, Mark A Schembri
Objectives: Polymyxins remain one of the last-resort drugs to treat infections caused by MDR Gram-negative pathogens. Here, we determined the mechanisms by which chromosomally encoded resistance to colistin and polymyxin B can arise in the MDR uropathogenic Escherichia coli ST131 reference strain EC958. Methods: Two complementary approaches, saturated transposon mutagenesis and spontaneous mutation induction with high concentrations of colistin and polymyxin B, were employed to select for mutations associated with resistance to polymyxins...
October 1, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29066548/genome-wide-discovery-of-genes-required-for-capsule-production-by-uropathogenic-escherichia-coli
#17
Kelvin G K Goh, Minh-Duy Phan, Brian M Forde, Teik Min Chong, Wai-Fong Yin, Kok-Gan Chan, Glen C Ulett, Matthew J Sweet, Scott A Beatson, Mark A Schembri
Uropathogenic Escherichia coli (UPEC) is a major cause of urinary tract and bloodstream infections and possesses an array of virulence factors for colonization, survival, and persistence. One such factor is the polysaccharide K capsule. Among the different K capsule types, the K1 serotype is strongly associated with UPEC infection. In this study, we completely sequenced the K1 UPEC urosepsis strain PA45B and employed a novel combination of a lytic K1 capsule-specific phage, saturated Tn5 transposon mutagenesis, and high-throughput transposon-directed insertion site sequencing (TraDIS) to identify the complement of genes required for capsule production...
October 24, 2017: MBio
https://www.readbyqxmd.com/read/29059366/sbcddb-sleeping-beauty-cancer-driver-database-for-gene-discovery-in-mouse-models-of-human-cancers
#18
Justin Y Newberg, Karen M Mann, Michael B Mann, Nancy A Jenkins, Neal G Copeland
Large-scale oncogenomic studies have identified few frequently mutated cancer drivers and hundreds of infrequently mutated drivers. Defining the biological context for rare driving events is fundamentally important to increasing our understanding of the druggable pathways in cancer. Sleeping Beauty (SB) insertional mutagenesis is a powerful gene discovery tool used to model human cancers in mice. Our lab and others have published a number of studies that identify cancer drivers from these models using various statistical and computational approaches...
October 20, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29032547/genome-wide-mutagenesis-in-borrelia-burgdorferi
#19
Tao Lin, Lihui Gao
Signature-tagged mutagenesis (STM) is a functional genomics approach to identify bacterial virulence determinants and virulence factors by simultaneously screening multiple mutants in a single host animal, and has been utilized extensively for the study of bacterial pathogenesis, host-pathogen interactions, and spirochete and tick biology. The signature-tagged transposon mutagenesis has been developed to investigate virulence determinants and pathogenesis of Borrelia burgdorferi. Mutants in genes important in virulence are identified by negative selection in which the mutants fail to colonize or disseminate in the animal host and tick vector...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29026163/revealing-the-saline-adaptation-strategies-of-the-halophilic-bacterium-halomonas-beimenensis-through-high-throughput-omics-and-transposon-mutagenesis-approaches
#20
Yan-Huey Chen, Chia-Wei Lu, Yuan-Tay Shyu, Shih-Shun Lin
Studies on the halotolerance of bacteria are attractive to the fermentation industry. However, a lack of sufficient genomic information has precluded an investigation of the halotolerance of Halomonas beimenensis. Here, we describe the molecular mechanisms of saline adaptation in H. beimenensis based on high-throughput omics and Tn5 transposon mutagenesis. The H. beimenensis genome is 4.05 Mbp and contains 3,807 genes, which were sequenced using short and long reads obtained via deep sequencing. Sixteen Tn5 mutants with a loss of halotolerance were identified...
October 12, 2017: Scientific Reports
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