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Letteratura scientifica selezionata sul tema "Flagella (Microbiology)"

Autore: Grafiati

Pubblicato: 4 giugno 2021

Ultima modifica: 3 marzo 2023

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Articoli di riviste sul tema "Flagella (Microbiology)"

1

Kanbe, Masaomi, Jin Yagasaki, Susanne Zehner, Michael Göttfert e Shin-Ichi Aizawa. "Characterization of Two Sets of Subpolar Flagella in Bradyrhizobium japonicum". Journal of Bacteriology 189, n. 3 (10 novembre 2006): 1083–89. http://dx.doi.org/10.1128/jb.01405-06.

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ABSTRACT Bradyrhizobium japonicum is one of the soil bacteria that form nodules on soybean roots. The cell has two sets of flagellar systems, one thick flagellum and a few thin flagella, uniquely growing at subpolar positions. The thick flagellum appears to be semicoiled in morphology, and the thin flagella were in a tight-curly form as observed by dark-field microscopy. Flagellin genes were identified from the amino acid sequence of each flagellin. Flagellar genes for the thick flagellum are scattered into several clusters on the genome, while those genes for the thin flagellum are compactly organized in one cluster. Both types of flagella are powered by proton-driven motors. The swimming propulsion is supplied mainly by the thick flagellum. B. japonicum flagellar systems resemble the polar-lateral flagellar systems of Vibrio species but differ in several aspects.

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2

Campodónico, Victoria L., Nicolás J. Llosa, Martha Grout, Gerd Döring, Tomás Maira-Litrán e Gerald B. Pier. "Evaluation of Flagella and Flagellin of Pseudomonas aeruginosa as Vaccines". Infection and Immunity 78, n. 2 (7 dicembre 2009): 746–55. http://dx.doi.org/10.1128/iai.00806-09.

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ABSTRACT Pseudomonas aeruginosa is a serious pathogen in hospitalized, immunocompromised, and cystic fibrosis (CF) patients. P. aeruginosa is motile via a single polar flagellum made of polymerized flagellin proteins differentiated into two major serotypes: a and b. Antibodies to flagella delay onset of infection in CF patients, but whether immunity to polymeric flagella and that to monomeric flagellin are comparable has not been addressed, nor has the question of whether such antibodies might negatively impact Toll-like receptor 5 (TLR5) activation, an important component of innate immunity to P. aeruginosa. We compared immunization with flagella and that with flagellin for in vitro effects on motility, opsonic killing, and protective efficacy using a mouse pneumonia model. Antibodies to flagella were superior to antibodies to flagellin at inhibiting motility, promoting opsonic killing, and mediating protection against P. aeruginosa pneumonia in mice. Protection against the flagellar type strains PAK and PA01 was maximal, but it was only marginal against motile clinical isolates from flagellum-immunized CF patients who nonetheless became colonized with P. aeruginosa. Purified flagellin was a more potent activator of TLR5 than were flagella and also elicited higher TLR5-neutralizing antibodies than did immunization with flagella. Antibody to type a but not type b flagella or flagellin inhibited TLR5 activation by whole bacterial cells. Overall, intact flagella appear to be superior for generating immunity to P. aeruginosa, and flagellin monomers might induce antibodies capable of neutralizing innate immunity due to TLR5 activation, but solid immunity to P. aeruginosa based on flagellar antigens may require additional components beyond type a and type b proteins from prototype strains.

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3

Marathe, Sandhya Amol, Arjun Balakrishnan, Vidya Devi Negi, Deepika Sakorey, Nagasuma Chandra e Dipshikha Chakravortty. "Curcumin Reduces the Motility of Salmonella enterica Serovar Typhimurium by Binding to the Flagella, Thereby Leading to Flagellar Fragility and Shedding". Journal of Bacteriology 198, n. 13 (18 aprile 2016): 1798–811. http://dx.doi.org/10.1128/jb.00092-16.

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ABSTRACTOne of the important virulence properties of the pathogen is its ability to travel to a favorable environment, cross the viscous mucus barrier (intestinal barrier for enteric pathogens), and reach the epithelia to initiate pathogenesis with the help of an appendage, like flagella. Nonetheless, flagella can act as an “Achilles heel,” revealing the pathogen's presence to the host through the stimulation of innate and adaptive immune responses. We assessed whether curcumin, a dietary polyphenol, could alter the motility ofSalmonella, a foodborne pathogen. It reduced the motility ofSalmonella entericaserovar Typhimurium by shortening the length of the flagellar filament (from ∼8 μm to ∼5 μm) and decreasing its density (4 or 5 flagella/bacterium instead of 8 or 9 flagella/bacterium). Upon curcumin treatment, the percentage of flagellated bacteria declined from ∼84% to 59%. However, no change was detected in the expression of the flagellin gene and protein. A fluorescence binding assay demonstrated binding of curcumin to the flagellar filament. This might make the filament fragile, breaking it into smaller fragments. Computational analysis predicted the binding of curcumin, its analogues, and its degraded products to a flagellin molecule at an interface between domains D1 and D2. Site-directed mutagenesis and a fluorescence binding assay confirmed the binding of curcumin to flagellin at residues ASN120, ASP123, ASN163, SER164, ASN173, and GLN175.IMPORTANCEThis work, to our knowledge the first report of its kind, examines how curcumin targets flagellar density and affects the pathogenesis of bacteria. We found that curcumin does not affect any of the flagellar synthesis genes. Instead, it binds to the flagellum and makes it fragile. It increases the torsional stress on the flagellar filament that then breaks, leaving fewer flagella around the bacteria. Flagella, which are crucial ligands for Toll-like receptor 5, are some of the most important appendages ofSalmonella. Curcumin is an important component of turmeric, which is a major spice used in Asian cooking. The loss of flagella can, in turn, change the pathogenesis of bacteria, making them more robust and fit in the host.

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4

Iino, T., Tomoko Oguchi e T. Kuroiwa. "Polymorphism in a Flagellar-shape Mutant of Salmonella typhimurium". Microbiology 81, n. 1 (1 gennaio 2000): 37–45. http://dx.doi.org/10.1099/00221287-81-1-37.

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A flagellar-shape mutant, designated ‘polymorphous’, was isolated from a normal flagella strain of Salmonella typhimurium. The mutant produces normal flagella in phase 1 and polymorphous flagella in phase 2. The polymorphous flagella are either straight or possess one of the four distinct wave-forms, namely M, S, N or C, when observed with an electron microscope after negative staining with phosphotungstic acid or uranyl acetate. Conversions between the four wave-forms were found to be brought about mainly by a change in the degree of twisting of longitudinal strands around the axis of a flagella filament, without marked change in the relative lengths of the outermost and innermost strands. The major fraction of the polymorphous mutant flagella showed the N-form under any conditions of specimen preparation. The remaining four forms appeared as minor fractions in various proportions. Specimens fixed with formalin showed less pronounced polymorphism than unfixed ones. Negative staining with uranyl acetate was more effective than with phosphotungstic acid for observing polymorphism. Even though more than one form appeared among the polymorphous flagella, each individual flagellum comprised a single form except for a rare coexistence of S and N. The same form of flagella tended to coexist in a bacterium in a heteromorphously flagellated cell population. It was concluded that the conformation and arrangement of the flagellin molecules responsible for wave-form result from strong mutual interactions between the neighbouring molecules along the flagellar filaments and also, to a lesser extent, between the neighbouring filaments in a flagellar bundle, as well as being influenced by the physico-chemical environment.

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5

Bardy, Sonia L., Takahisa Mori, Kaoru Komoriya, Shin-Ichi Aizawa e Ken F. Jarrell. "Identification and Localization of Flagellins FlaA and FlaB3 within Flagella of Methanococcus voltae". Journal of Bacteriology 184, n. 19 (1 ottobre 2002): 5223–33. http://dx.doi.org/10.1128/jb.184.19.5223-5233.2002.

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ABSTRACT Methanococcus voltae possesses four flagellin genes, two of which (flaB1 and flaB2) have previously been reported to encode major components of the flagellar filament. The remaining two flagellin genes, flaA and flaB3, are transcribed at lower levels, and the corresponding proteins remained undetected prior to this work. Electron microscopy examination of flagella isolated by detergent extraction of whole cells revealed a curved, hook-like region of varying length at the end of a long filament. Enrichment of the curved region of the flagella resulted in the identification of FlaB3 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and N-terminal sequencing, and the localization of this flagellin to the cell-proximal portion of the flagellum was confirmed through immunoblotting and immunoelectron microscopy with FlaB3-specific antibodies, indicating that FlaB3 likely composes the curved portion of the flagella. This could represent a unique case of a flagellin performing the role of the bacterial hook protein. FlaA-specific antibodies were used in immunoblotting to determine that FlaA is found throughout the flagellar filament. M. voltae cells were transformed with a modified flaA gene containing a hemagglutinin (HA) tag introduced into the variable region. Transformants that had replaced the wild-type copy of the flaA gene with the HA-tagged version incorporated the HA-tagged version of FlaA into flagella which appeared normal by electron microscopy.

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6

del Campo, Ana Martínez, Teresa Ballado, Javier de la Mora, Sebastian Poggio, Laura Camarena e Georges Dreyfus. "Chemotactic Control of the Two Flagellar Systems of Rhodobacter sphaeroides Is Mediated by Different Sets of CheY and FliM Proteins". Journal of Bacteriology 189, n. 22 (21 settembre 2007): 8397–401. http://dx.doi.org/10.1128/jb.00730-07.

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ABSTRACT Rhodobacter sphaeroides expresses two different flagellar systems, a subpolar flagellum (fla1) and multiple polar flagella (fla2). These structures are encoded by different sets of flagellar genes. The chemotactic control of the subpolar flagellum (fla1) is mediated by three of the six different CheY proteins (CheY6, CheY4, or CheY3). We show evidence that CheY1, CheY2, and CheY5 control the chemotactic behavior mediated by fla2 flagella and that RSP6099 encodes the fla2 FliM protein.

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7

Rabaan, Ali A., Ioannis Gryllos, Juan M. Tomás e Jonathan G. Shaw. "Motility and the Polar Flagellum Are Required for Aeromonas caviae Adherence to HEp-2 Cells". Infection and Immunity 69, n. 7 (1 luglio 2001): 4257–67. http://dx.doi.org/10.1128/iai.69.7.4257-4267.2001.

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ABSTRACT Aeromonas caviae is increasingly being recognized as a cause of gastroenteritis, especially among the young. The adherence of aeromonads to human epithelial cells in vitro has been correlated with enteropathogenicity, but the mechanism is far from well understood. Initial investigations demonstrated that adherence of A. caviae to HEp-2 cells was significantly reduced by either pretreating bacterial cells with an antipolar flagellin antibody or by pretreating HEp-2 cells with partially purified flagella. To precisely define the role of the polar flagellum in aeromonad adherence, we isolated the A. caviae polar flagellin locus and identified five polar flagellar genes, in the order flaA, flaB, flaG, flaH, and flaJ. Each gene was inactivated using a kanamycin resistance cartridge that ensures the transcription of downstream genes, and the resulting mutants were tested for motility, flagellin expression, and adherence to HEp-2 cells. N-terminal amino acid sequencing, mutant analysis, and Western blotting demonstrated that A. caviae has a complex flagellum filament composed of two flagellin subunits encoded by flaAand flaB. The predicted molecular mass of both flagellins was ∼31,700 Da; however, their molecular mass estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was ∼35,500 Da. This aberrant migration was thought to be due to their glycosylation, since the proteins were reactive in glycosyl group detection assays. Single mutations in either flaA orflaB did not result in loss of flagella but did result in decreased motility and adherence by approximately 50%. Mutation offlaH, flaJ, or both flagellin genes resulted in the complete loss of motility, flagellin expression, and adherence. However, mutation of flaG did not affect motility but did significantly reduce the level of adherence. Centrifugation of the flagellate mutants (flaA, flaB, and flaG) onto the cell monolayers did not increase adherence, whereas centrifugation of the aflagellate mutants (flaH, flaJ, and flaA flaB) increased adherence slightly. We conclude that maximum adherence of A. caviae to human epithelial cells in vitro requires motility and optimal flagellar function.

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Kirov, Sylvia M., Bronwen C. Tassell, Annalese B. T. Semmler, Lisa A. O’Donovan, Ali A. Rabaan e Jonathan G. Shaw. "Lateral Flagella and Swarming Motility in Aeromonas Species". Journal of Bacteriology 184, n. 2 (15 gennaio 2002): 547–55. http://dx.doi.org/10.1128/jb.184.2.547-555.2002.

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ABSTRACT Swarming motility, a flagellum-dependent behavior that allows bacteria to move over solid surfaces, has been implicated in biofilm formation and bacterial virulence. In this study, light and electron microscopic analyses and genetic and functional investigations have shown that at least 50% of Aeromonas isolates from the species most commonly associated with diarrheal illness produce lateral flagella which mediate swarming motility. Aeromonas lateral flagella were optimally produced when bacteria were grown on solid medium for ≈8 h. Transmission and thin-section electron microscopy confirmed that these flagella do not possess a sheath structure. Southern analysis of Aeromonas reference strains and strains of mesophilic species (n = 84, varied sources and geographic regions) with a probe designed to detect lateral flagellin genes (lafA1 and lafA2) showed there was no marked species association of laf distribution. Approximately 50% of these strains hybridized strongly with the probe, in good agreement with the expression studies. We established a reproducible swarming assay (0.5% Eiken agar in Difco broth, 30°C) for Aeromonas spp. The laf-positive strains exhibited vigorous swarming motility, whereas laf-negative strains grew but showed no movement from the inoculation site. Light and scanning electron microscopic investigations revealed that lateral flagella formed bacterium-bacterium linkages on the agar surface. Strains of an Aeromonas caviae isolate in which lateral flagellum expression was abrogated by specific mutations in flagellar genes did not swarm, proving conclusively that lateral flagella are required for the surface movement. Whether lateral flagella and swarming motility contribute to Aeromonas intestinal colonization and virulence remains to be determined.

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Canals, Rocío, Maria Altarriba, Silvia Vilches, Gavin Horsburgh, Jonathan G. Shaw, Juan M. Tomás e Susana Merino. "Analysis of the Lateral Flagellar Gene System of Aeromonas hydrophila AH-3". Journal of Bacteriology 188, n. 3 (1 febbraio 2006): 852–62. http://dx.doi.org/10.1128/jb.188.3.852-862.2006.

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ABSTRACT Mesophilic Aeromonas strains express a polar flagellum in all culture conditions, and certain strains produce lateral flagella on semisolid media or on surfaces. Although Aeromonas lateral flagella have been described as a colonization factor, little is known about their organization and expression. Here we characterized the complete lateral flagellar gene cluster of Aeromonas hydrophila AH-3 containing 38 genes, 9 of which (lafA-U) have been reported previously. Among the flgL L and lafA structural genes we found a modification accessory factor gene (maf-5) that is involved in formation of lateral flagella; this is the first time that such a gene has been described for lateral flagellar gene systems. All Aeromonas lateral flagellar genes were located in a unique chromosomal region, in contrast to Vibrio parahaemolyticus, in which the analogous genes are distributed in two different chromosomal regions. In A. hydrophila mutations in flhA L, lafK, fliJ L, flgN L, flgE L, and maf-5 resulted in a loss of lateral flagella and reductions in adherence and biofilm formation, but they did not affect polar flagellum synthesis. Furthermore, we also cloned and sequenced the A. hydrophila AH-3 alternative sigma factor σ54 (rpoN); mutation of this factor suggested that it is involved in expression of both types of flagella.

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10

Yen, Jiun Y., Katherine M. Broadway e Birgit E. Scharf. "Minimum Requirements of Flagellation and Motility for Infection of Agrobacterium sp. Strain H13-3 by Flagellotropic Bacteriophage 7-7-1". Applied and Environmental Microbiology 78, n. 20 (3 agosto 2012): 7216–22. http://dx.doi.org/10.1128/aem.01082-12.

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ABSTRACTThe flagellotropic phage 7-7-1 specifically adsorbs toAgrobacteriumsp. strain H13-3 (formerlyRhizobium lupiniH13-3) flagella for efficient host infection. TheAgrobacteriumsp. H13-3 flagellum is complex and consists of three flagellin proteins: the primary flagellin FlaA, which is essential for motility, and the secondary flagellins FlaB and FlaD, which have minor functions in motility. Using quantitative infectivity assays, we showed that absence of FlaD had no effect on phage infection, while absence of FlaB resulted in a 2.5-fold increase in infectivity. AflaAdeletion strain, which produces straight and severely truncated flagella, experienced a significantly reduced infectivity, similar to that of aflaB flaDstrain, which produces a low number of straight flagella. A strain lacking all three flagellin genes is phage resistant. In addition to flagellation, flagellar rotation is required for infection. A strain that is nonmotile due to an in-frame deletion in the gene encoding the motor component MotA is resistant to phage infection. We also generated two strains with point mutations in themotAgene resulting in replacement of the conserved charged residue Glu98, which is important for modulation of rotary speed. A change to the neutral Gln caused the flagellar motor to rotate at a constant high speed, allowing a 2.2-fold-enhanced infectivity. A change to the positively charged Lys caused a jiggly motility phenotype with very slow flagellar rotation, which significantly reduced the efficiency of infection. In conclusion, flagellar number and length, as well as speed of flagellar rotation, are important determinants for infection by phage 7-7-1.

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Più fonti

Tesi sul tema "Flagella (Microbiology)"

1

Wan, Yixin. "Modulation and synchronization of eukaryotic flagella". Thesis, University of Cambridge, 2014. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.708434.

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Moura, Cláudia de. "Identificação de novos antígenos flagelares e variação de fase em amostras de Escherichia coli isoladas de animais e alimentos". [s.n.], 2010. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317446.

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Orientador: Domingos da Silva Leite
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Escherichia coli é um membro comensal da microbiota de animais, porém podem causar doenças desde diarréias até sepses. A caracterização dos seus antígenos de superfície O (somático) e H (flagelar) auxilia na determinação de linhagens patogênicas dentro da espécie. Contudo, algumas bactérias não expressam flagelo in vitro, demonstrado que a amplificação do gene fliC, a análise dos fragmentos de polimorfismo (PCR-RFLP) e sequenciamento podem ser utilizadas para identificação dos antígenos H, em substituição à sorologia convencional. Até meados de 1980, pensava-se que, diferentemente da Salmonella, E. coli possui um único gene para expressão de flagelina (fliC), mas algumas amostras podem conter genes para expressão de flagelina flkA, fllA, flmA, flnA e fljA (repressor de fliC). Em nosso trabalho, analisamos 31 amostras de E. coli isolados de animais e alimentos que apresentavam o fenótipo HNT em ensaios de sorologia. Utilizamos PCR-RFLP e sequenciamento para descrever novos genes para flagelina, da qual foram obtidos antissoros. Identificamos por PCR e sequenciamento os genes responsáveis pela variação de fase fljA, flkA e flmA, realizamos experimentos de motilidade para determinar a variação de fase flagelar e detectar a expressão dos genes através de RT-PCR. Dezessete amostras tiveram seus antígenos H caracterizados, sendo nove caracterizadas por PCR-RFLP: H2 (duas amostras) H16 (duas amostras), H34 (três amostras), H33 (uma amostra) e H38 (uma amostra). Na análise de sequenciamento identificamos duas amostras portadoras do gene fliCh25, duas amostras fliCh7 e uma amostra apresentando fliCh32. Três novos genes para flagelina foram descritos: fliCh2', fliC4c, fliC40c. Identificamos o gene fljA em duas amostras HNT (3C e 4C) e na amostra padrão H35. O gene das amostras HNT apresentaram homologia ao fljA de Salmonella enterica, cuja variação de fase é bem estabelecida. As amostras padrão H11, H35, H40 e H47, bem como as amostras HNT 3C e 4C foram positivas para o gene flmA. As amostras padrão H3 e H53 são portadoras do gene flkA, contudo apenas a amostra H53 apresentou fljA. A amostra H54 é portadora de fljA e flmA. Nenhuma amostra H padrão mostrou variação de fase, diferentemente da literatura, sugerindo a perda da capacidade de variar a fase flagelar. A amostra 4C mostrou variação de fase positiva quando induzida em meios de cultura contendo antissoros anti-H48, anti-H54 e anti-H4C. Do mesmo modo, a detecção dos RNAm em diferentes condições de cultura confirmou a variação de fase. Como resultado um esquema de identificação para detecção de grupos de antígenos H e identificação de fliC foi testado. A técnica de fliC-RFLP provou ser eficiente e rápida, auxiliando a sorologia clássica para detecção de antígenos H de E. coli. Um modelo geral de variação de fase da amostra 4C é expresso por fliCoff + flmAon ? fliCon + flmAoff. Além disso, nós verificamos que a amostra 4C apresenta um gene novo para expressão de flagelina. Este trabalho é pioneiro em relação à variação de fase flagelar, demonstrando uma nova associação entre os antígenos H48 e H54
Abstract: Escherichia coli are a species of microflora, and characterization of the cell surface lipopolysaccharide O antigen and the flagellar H antigen allow the grouping of pathogenic clones within this species. Moreover, some bacteria in vitro do not obtain to express its flagella, demonstrated that PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing analysis has been used for the identification of these antigens, in substitution of traditional serology. Moreover, until middle of years 80, are believed, differently of the Salmonella, E. coli possesss an only gene for flagelin expression (fliC), but some s/strains can contain genes for flagellin expression flkA, fllA, flmA, flnA and fljA (repressor of fliC). In this work, we analyzed 31 strains of E. coli isolated from animals and foods that presented HNT phenotype in serology assays. We use PCR-RFLP and sequencing to describe new genes for flagellin, of which antiserum were obtained. We identify for PCR and sequencing the genes for phase variation fljA, flkA and flmA, we carry through motility experiments to determine the flagellar phase variation and to detect the expression of the genes (RNAm) through RT-PCR. Seventeen strains had had its H antigen characterized and nine of then were characterized for PCR-RFLP: H2 (two strains) H16 (two strains), H34 (three strains), H33 (one strain) and H38 (one strain). Through sequencing analysis we identify to two carrying strains of the gene fliCh25, two strains fliCh7 and one strain presenting fliCh32. Three new genes for flagellin had been described: fliCh2', fliC4c, fliC40c. Using PCR and sequencing, we identify fljA gene in two strains HNT (3C and 4C) and in the H35 control strain. The HNT genes showed homology to fljA of Salmonella enterica, whose variation of phase well is established. The control strains H11, H35, H40 and H47, as well as HNT 3C and 4C strains were positive for flmA gene. The control strains H3 and H53 are carrying of flkA gene, however only the H53 strain presented fljA. The H54 control strain is carrying of fljA and flmA. No H control strain showed phase variation, differently of literature, suggesting the loss of the capacity to flagellar phase variation. The 4C strain showed positive phase variation when cultured with antiserum anti-H48, anti-H54 and anti-H4C. In a similar way, the detention of RNAm in different conditions of culture confirmed the phase variation. As a result, an identification scheme was tested to deduce H antigen groups and new genes of fliC. The fliCRFLP technique proved to be faster than classic serotyping for the deduction of the E. coli H antigen, characterizing the antigens with few days and indicating new putative genes. Thus, a general model for flagellar phase variation in 4C strain can be expressed as fliCoff + flmAon ? fliCon + flmAoff. In addition, we found that strains 3C and 4C express unidentified flagellin antigens. This is the first report of flagellar phase variation in wild E. coli strains. We have also provided evidence that strain 4C, identified here for the first time, expresses three flagellar antigens, H48, H54 and a previously unidentified flagellin
Doutorado
Microbiologia
Doutor em Genetica e Biologia Molecular

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3

Pacheco, Sophia A. "Identification of Campylobacter jejuni secreted proteins". Pullman, Wash. : Washington State University, 2010. http://www.dissertations.wsu.edu/Thesis/Spring2010/s_pacheco_021610.pdf.

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4

Wang, Xiaoling. "Mechanical analysis and free energy construction of phase transition in bacterial flagellar filaments /". View abstract or full-text, 2006. http://library.ust.hk/cgi/db/thesis.pl?MECH%202006%20WANGX.

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5

Steager, Edward Brian Kim MinJun. "Actuation and control of microfabricated structures using flagellated bacteria /". Philadelphia, Pa. : Drexel University, 2009. http://hdl.handle.net/1860/3132.

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6

Tiba, Monique Ribeiro. "Identificação de novos antigenos flagelares de Escherichia coli de origem humana". [s.n.], 2009. http://repositorio.unicamp.br/jspui/handle/REPOSIP/316681.

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Orientador: Domingos da Silva Leite
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Escherichia coli tem sido isolada, com certa freqüência, apresentando antígenos flagelares (H) que não são reconhecidos por nenhum dos anti-soros disponibilizado pelo mais importante centro de referência de E. coli, The International Escherichia and Klebsiella Centre (WHO) do Statens Serum Institut, Copenhague, Dinamarca. Atualmente são reconhecidos 53 antígenos "H" e, nos últimos 29 anos, nenhuma modificação ocorreu na lista dos antígenos flagelares associados à Escherichia coli. Isto posto, os objetivos deste trabalho foram identificar os antígenos flagelares das cepas de E. coli que expressam H não tipável (HNT) e que apresentam fatores de virulência associados à diferentes enteropatias. Esta identificação foi realizada inicialmente, pela reação em cadeia da polimerase (PCR) do gene fliC, responsável pela proteína flagelina, das 53 amostras padrões para os antígenos H e das 20 amostras HNT (H não-tipável). Em seguida, os amplicons foram digeridos por enzimas de restrição e daquelas amostras que apresentaram perfis de restrições distintos daqueles observados para as amostras padrões de antígeno H, foram produzidos soros em coelhos. Foram realizados testes de titulação frente aos 53 antígenos padrões, frente ao antígeno homólogo e frente aos antígenos das amostras HNT. As seqüência gênicas das amostras HNT, obtidas na reação de sequenciamento, foram comparadas aos diferentes genes de fliC armazenados no banco de dados do "National Center for Biotecnology Information" (NCBI) através do sistema BLAST, e o programa ClustalW foi utilizado para alinhamento das seqüências. Os resultados demonstraram que estas amostras apresentaram similaridade com antígenos padrões, entretanto, elas não possuem a mesma seqüência nucleotídica e também não reagiram fenotipicamente com o anti-soro esperado. Os dados obtidos permitem concluir que no conjunto de amostras estudado, treze amostras apresentaram antígeno flagelar diferente daqueles já descritos na literatura, quando utilizado as técnicas de PCR e/ou sorologia.
Abstract: Escherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Centre (WHO) of the Statens Serum Institute, Copenhagen, Denmark. Are currently recognized 53 H antigens and in the last 29 years, no change occurred in the list of flagellar antigens associated with Escherichia coli. The objectives of this study were to identify the flagellar antigens of E. coli that do not express non-typeable H antigens and presenting the virulence factors associated with different diseases. This identification was performed initially by gene amplification of the fliC, (flagellin protein) by the polymerase chain reaction (PCR) in all 53 standards E. coli strains for the H antigens and 20 non-typeable H-antigens E. coli strains, being then, the amplicons were digested by restriction enzymes. Anti-sera were produced in rabbits, those strains that showed different restriction profiles of these patterns observed for the nontypeable H antigens E. coli strains. Agglutination testes were carried out against the 53 antigens standards, against the homologous antigen and H antigens of the non-typeable strains. DNA sequences were compared to different fliC genes stored in the database of the National Center for Biotecnology Information (NCBI) through the BLAST, and ClustalW program was used to align the sequences. The results showed that although these strains have homology with a standard H-antigen, they do not have the same nucleotide sequence and did not phenotypically reacted with the antiserum expected. The data obtained showed that thirteen strains had a different H antigen those already described in the literature when used the techniques of PCR-RFLP and/or serology.
Doutorado
Microbiologia
Doutor em Genetica e Biologia Molecular

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Boateng, Lindsy R. "Determining the binding partners of PKA in the axoneme of Chlamydomonas reinhardtii flagella". Connect to online version, 2009. http://minds.wisconsin.edu/handle/1793/37450.

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Schreiber, Maria Fernanda. "Studying the host transcriptome and the role of flagella in infections mediated by Salmonella and other pathogens". Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610270.

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Marshall, Joanna M. "The O-Antigen Capsule of Salmonella Typhimurium in Acute and Chronic Infection". The Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1385998769.

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Azevedo, Fátima da Piedade de Melo. "Inserção de epitopo heterólogo em diferentes regiões de flagelina bacteriana: influência na função flagelar e imunogenicidade". Universidade de São Paulo, 1997. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-09112015-150421/.

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Abstract (sommario):

Uma das estratégias mais promissoras para a biotecnologia de vacinas é o desenvolvimento de linhagens precisamente atenuadas, e que possam ser usadas como carregadoras de antígenos heterólogos. Mutantes de Salmonella Typhimurium têm sido extensivamente utilizados com essa fmalidade. A flagelina, monômero constituinte do filamento flagelar, vem sendo empregada como carregadora de antígenos heterólogos, inseridos na região central, hipervariável (região IV). Inserções nessa região são freqüentemente funcionais, e levam à exposição do epitopo na superfície do filamento. O presente trabalho explora o potencial de outras regiões da molécula para a inserção de epitopos. Nós inserimos a mesma seqüência usada anteriomente (epitopo da proteína M de S. pyogenes, Tipo 5) em regiões com diferentes níveis de homologia (III e VI), e em região totalmente conservada (VIII). Também foram feitas inserções duplas em regiões que se mostraram toleráveis (III e IV; IV e VI). Todas as proteínas híbridas foram sintetizadas pela Salmonella, como demonstrado em imunoblots, usando anticorpo contra a flagelina e contra o peptídeo. Todas as regiões, exceto a VIII, aceitaram a inserção sem perda de motilidade, apesar de, em alguns casos, ela ter sido extremamente reduzida. A imunogenicidade foi avaliada pela imunização de camundongos com bactérias vivas, inativadas ou, quando possível, flagelina purificada. Os resultados foram similares aos descritos na literatura para inserções envolvendo a região IV, obtendo-se um elevado título de anticorpos contra flagelina. Um baixo nível de anticorpo contra o peptídeo também foi detectado para todas as novas linhagens testadas. Nossos resultados com imunização de bactérias vivas sugerem uma resposta levemente melhor ao peptídeo quando duas cópias estão presentes, mas os dados não são conclusivos.
One of the most promising strategies for the biotechnology of vaccines is the development of precisely attenuated strains, which could be used as carriers of heterologous antigens. Mutants of Salmonella Typhimurium have been extensively explored to this effect, since the infection ofmice by S. Typhimurium mimics the infection of humans by S. Typhi, and the genetics of the species is extremely well known, making it easy the obtention of defined mutants with reduced pathogenicity. Mutants with auxotrofy in genes of the aromatic pathway are particularly attractive, since they need PABA and DHB to grow, and these compounds are unavailable in mammalian tissues. Flagellin, the monomer which constitutes the flagellar filament, has been used as a carrier for heterologous epitopes, inserted in a central, hypervariable region (region IV). Insertions in this region are often functional, and lead to exposition of the epitope at the filament\' s surface. The present work explored the potential of the other regions ofthe molecule for the insertion of epitopes. We inserted the same reporter sequence (MS epitope from S. pyogenes M protein) in regions with different levels of homology (III and VI), and totally conserved (VIII). We also made double insertions in regions shown to be permissive (III and IV; IV and VI). All hybrid proteins were synthesized by Salmonella, as demonstrated by immunoblots using antibody against flagellin and against the synthetic peptide. All regions, except the highly conserved region VIII, accepted the insertions without loss of motility, albeit, in some cases, motility was seriously reduced. Immunogenicity of the hydrids was evaluated by immunization with live bacteria, killed bacteria, and purified flagellin (when possible). Results obtained with the new constructs were similar to the ones published for insertions involving region IV, in the sense that antibody titers to the carrier protein were very high. A low level of antibody to the inserted peptide was also detected in all groups of animals. Our results with live immunization suggest a slightly better response to the peptide when two copies are present, but the data are not conclusive.

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Libri sul tema "Flagella (Microbiology)"

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1953-, Jarrell Kenneth F., a cura di. Pili and flagella: Current research and future trends. Norfolk, UK: Caister Academic Press, 2009.

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1953-, Jarrell Kenneth F., a cura di. Pili and flagella: Current research and future trends. Norfolk, UK: Caister Academic Press, 2009.

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A, Bloodgood Robert, a cura di. Ciliary and flagellar membranes. New York: Plenum Press, 1990.

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Prüss, Birgit M. Global regulatory networks in enteric bacteria. Kerala, India: Research Signpost, 2005.

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Murase, Masatoshi. Dynamics of cellular motility. Chichester: J. Wiley & Sons, 1992.

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Murase, Mosatoshi. Dynamics of cellular motility. Chichester [England]: Wiley, 1992.

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Cilia: Model organisms and intraflagellar transport. Burlington, MA: Elsevier Academic Press, 2009.

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Atkins, Michael S. Assessment of flagellate diversity at deep-sea hydrothermal vents using the combined approach of culture-dependent and culture-independent methods. Cambridge, Mass: Massachusetts Institute of Technology, 2000.

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Flagella and Cilia: Types, Structure and Functions. Nova Science Publishers, Incorporated, 2018.

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Aizawa, Shin-Ichi. Flagellar World: Electron Microscopic Images of Bacterial Flagella and Related Surface Structures. Elsevier Science & Technology Books, 2013.

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Capitoli di libri sul tema "Flagella (Microbiology)"

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Buisson, Johanna, e Philippe Bastin. "Flagellum Structure and Function in Trypanosomes". In Microbiology Monographs, 63–86. Berlin, Heidelberg: Springer Berlin Heidelberg, 2010. http://dx.doi.org/10.1007/978-3-642-12863-9_3.

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McKean, Paul G., e Keith Gull. "The Flagellar Pocket of Trypanosomatids: A Critical Feature for Cell Morphogenesis and Pathogenicity". In Microbiology Monographs, 87–113. Berlin, Heidelberg: Springer Berlin Heidelberg, 2010. http://dx.doi.org/10.1007/978-3-642-12863-9_4.

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Schell, Ursula, Sylvia Simon e Hubert Hilbi. "Inflammasome Recognition and Regulation of the Legionella Flagellum". In Current Topics in Microbiology and Immunology, 161–81. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-41171-2_8.

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Givaudan, Alain, e Anne Lanois. "Flagellar Regulation and Virulence in the Entomopathogenic Bacteria—Xenorhabdus nematophila and Photorhabdus luminescens". In Current Topics in Microbiology and Immunology, 39–51. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/82_2016_53.

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Smith, K. D., e A. Ozinsky. "Toll-Like Receptor-5 and the Innate Immune Response to Bacterial Flagellin". In Current Topics in Microbiology and Immunology, 93–108. Berlin, Heidelberg: Springer Berlin Heidelberg, 2002. http://dx.doi.org/10.1007/978-3-642-59430-4_6.

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Aizawa, Shin-Ichi. "Flagella". In Molecular Medical Microbiology, 125–46. Elsevier, 2015. http://dx.doi.org/10.1016/b978-0-12-397169-2.00007-x.

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AIZAWA, S. "Flagella". In Molecular Medical Microbiology, 155–75. Elsevier, 2002. http://dx.doi.org/10.1016/b978-012677530-3/50227-0.

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Aizawa, S. I. "Flagella, Prokaryotic". In Encyclopedia of Microbiology, 393–403. Elsevier, 2009. http://dx.doi.org/10.1016/b978-012373944-5.00047-x.

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Sockett, R. Elizabeth. "6.5 Characterizing Flagella and Motile Behavior". In Methods in Microbiology, 227–38. Elsevier, 1998. http://dx.doi.org/10.1016/s0580-9517(08)70286-1.

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Panjarathinam, R. "Class Mastigophora (Flagellate)". In Practical Medical Microbiology, 111. Jaypee Brothers Medical Publishers (P) Ltd., 2009. http://dx.doi.org/10.5005/jp/books/11099_22.

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Atti di convegni sul tema "Flagella (Microbiology)"

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Oliveira, B. H., M. R. Silva, C. J. M. Braga, L. M. Massis, L. C. S. Ferreira, M. E. Sbrogio-Almeida e M. Takagi. "Salmonella enterica Typhimurium: establishment of cultivation condition on shake flask and flagellin isolation strategy by using tangencial ultrafiltration". In Proceedings of the III International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld2009). WORLD SCIENTIFIC, 2010. http://dx.doi.org/10.1142/9789814322119_0075.

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