Gotowe bibliografie tematyczne / RRNA gene analysis

Gotowa bibliografia na temat „RRNA gene analysis”

Autor: Grafiati
Data publikacji: 4 czerwca 2021
Data aktualizacji: 20 lutego 2023

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Artykuły w czasopismach na temat "RRNA gene analysis"

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Liu, Junjie, Peng Li, Liuyang Lu, Lanfen Xie, Xiling Chen i Baizhong Zhang. "Selection and evaluation of potential reference genes for gene expression analysis in Avena fatua Linn". Plant Protection Science 55, No. 1 (20.11.2018): 61–71. http://dx.doi.org/10.17221/20/2018-pps.

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Eight commonly used candidate reference genes, 18S ribosomal RNA (rRNA) (18S), 28S rRNA (28S), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1 alpha (EF1α), ribosomal protein L7 (RPL7), Alpha-tubulin (α-TUB), and TATA box binding protein-associated factor (TBP), were evaluated under various experimental conditions to assess their suitability in different developmental stages, tissues and herbicide treatments in Avena fatua. The results indicated the most suitable reference genes for the different experimental conditions. For developmental stages, 28S and EF1α were the optimal reference genes, both EF1α and 28S were suitable for experiments of different tissues, whereas for herbicide treatments, GAPDH and ACT were suitable for normalizations of expression data. In addition, GAPDH and EF1α were the suitable reference genes.
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Gonzalez-y-Merchand, J. A., M. J. Colston i R. A. Cox. "Effects of Growth Conditions on Expression of Mycobacterial murA and tyrS Genes and Contributions of Their Transcripts to Precursor rRNA Synthesis". Journal of Bacteriology 181, nr 15 (1.08.1999): 4617–27. http://dx.doi.org/10.1128/jb.181.15.4617-4627.1999.

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ABSTRACT All mycobacteria studied to date have an rRNA operon, designatedrrnA, located downstream from a single copy of themurA gene, which encodes an enzyme (EC 2.5.1.7 ) important for peptidoglycan synthesis. The rrnA operon has a promoter, P1(A), located within the coding region of murA, near the 3′ end. Samples of RNA were isolated from Mycobacterium tuberculosis at different stages of the growth cycle and fromMycobacterium smegmatis grown under different conditions. RNase protection assays were used to investigate transcripts of bothmurA and rrnA. Transcription ofmurA was found to continue into the 16S rRNA gene, as ifmurA and rrnA form a hybrid (protein coding-rRNA coding) operon. During the growth of M. tuberculosis, the hybrid operon contributed approximately 2% to total pre-rRNA. Analysis of M. smegmatis RNA revealed that the level of murA RNA depended on the growth rate and that the patterns of expression during the growth cycle were different formurA and rrnA. M. smegmatis has a second rRNA operon, rrnB, located downstream from a single copy of the tyrS gene, encoding tyrosyl-tRNA synthetase. Transcription of tyrS was found to continue into the 16S rRNA gene rrnB. The hybrid tyrS-rrnB operon contributed 0.2 to 0.6% to rrnB transcripts. The pattern of tyrS expression during the growth cycle matched the pattern of rrnB expression, reflecting the essential role of TyrS and rRNA in protein biosynthesis.
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Yap, Wai Ho, Zhenshui Zhang i Yue Wang. "Distinct Types of rRNA Operons Exist in the Genome of the Actinomycete Thermomonospora chromogena and Evidence for Horizontal Transfer of an Entire rRNA Operon". Journal of Bacteriology 181, nr 17 (1.09.1999): 5201–9. http://dx.doi.org/10.1128/jb.181.17.5201-5209.1999.

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ABSTRACT We describe here the presence of two distinct types of rRNA operons in the genome of a thermophilic actinomycete Thermomonospora chromogena. The genome of T. chromogena contains six rRNA operons (rrn), of which four complete and two incomplete ones were cloned and sequenced. Comparative analysis revealed that the operon rrnB exhibits high levels of sequence variations to the other five nearly identical ones throughout the entire length of the operon. The coding sequences for the 16S and 23S rRNA genes differ by approximately 6 and 10%, respectively, between the two types of operons. Normal functionality ofrrnB is concluded on the basis of the nonrandom distribution of nucleotide substitutions, the presence of compensating nucleotide covariations, the preservation of secondary and tertiary rRNA structures, and the detection of correctly processed rRNAs in the cell. Comparative sequence analysis also revealed a close evolutionary relationship between rrnB operon of T. chromogena and rrnA operon of another thermophilic actinomycete Thermobispora bispora. We propose thatT. chromogena acquired rrnB operon fromT. bispora or a related organism via horizontal gene transfer.
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Menendez, M. C., M. J. Garcia, M. C. Navarro, J. A. Gonzalez-y-Merchand, S. Rivera-Gutierrez, L. Garcia-Sanchez i R. A. Cox. "Characterization of an rRNA Operon (rrnB) of Mycobacterium fortuitum and Other Mycobacterial Species: Implications for the Classification of Mycobacteria". Journal of Bacteriology 184, nr 4 (15.02.2002): 1078–88. http://dx.doi.org/10.1128/jb.184.4.1078-1088.2002.

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ABSTRACT Mycobacteria are thought to have either one or two rRNA operons per genome. All mycobacteria investigated to date have an operon, designated rrnA, located downstream from the murA gene. We report that Mycobacteriun fortuitum has a second rrn operon, designated rrnB, which is located downstream from the tyrS gene; tyrS is very close to the 3" end of a gene (3-mag) coding for 3-methylpurine-DNA-glycosylase. The second rrn operon of Mycobacterium smegmatis was shown to have a similar organization, namely, 5" 3-mag-tyrS-rrnB 3". The rrnB operon of M. fortuitum was found to have a single dedicated promoter. During exponential growth in a rich medium, the rrnB and rrnA operons were the major and minor contributors, respectively, to pre-rRNA synthesis. Genomic DNA was isolated from eight other fast-growing mycobacterial species. Samples were investigated by Southern blot analysis using probes for murA, tyrS, and 16S rRNA sequences. The results revealed that both rrnA and rrnB operons were present in each species. The results form the basis for a proposed new scheme for the classification of mycobacteria. The approach, which is phylogenetic in concept, is based on particular properties of the rrn operons of a cell, namely, the number per genome and a feature of 16S rRNA gene sequences.
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Prammananan, Therdsak, Peter Sander, Burkhard Springer i Erik C. Böttger. "RecA-Mediated Gene Conversion and Aminoglycoside Resistance in Strains Heterozygous for rRNA". Antimicrobial Agents and Chemotherapy 43, nr 3 (1.03.1999): 447–53. http://dx.doi.org/10.1128/aac.43.3.447.

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ABSTRACT Clinical resistance to aminoglycosides in general is due to enzymatic drug modification. Mutational alterations of the small ribosomal subunit rRNA have recently been found to mediate acquired resistance in bacterial pathogens in vivo. In this study we investigated the effect of 16S rRNA heterozygosity (wild-type [wt] and mutant [mut] operons at position 1408 [1408wt/1408mut]) on aminoglycoside resistance. Using an integrative vector, we introduced a single copy of a mutated rRNA operon (1408 A→G) into Mycobacterium smegmatis, which carries two chromosomal wild-type rRNA operons; the resultant transformants exhibited an aminoglycoside-sensitive phenotype. In contrast, introduction of the mutated rRNA operon into anM. smegmatis rrnB knockout strain carrying a single functional chromosomal wild-type rRNA operon resulted in aminoglycoside-resistant transformants. Subsequent analysis by DNA sequencing and RNase protection assays unexpectedly demonstrated a homozygous mutant genotype, rRNAmut/rRNAmut, in the resistant transformants. To investigate whether RecA-mediated gene conversion was responsible for the aminoglycoside-resistant phenotype in the rRNAwt/rRNAmut strains, recAmutant strains were generated by allelic exchange techniques. Transformation of the recA rrnB M. smegmatis mutant strains with an integrative vector expressing a mutated rRNA operon (Escherichia coli position 1408 A→G) resulted in transformants with an aminoglycoside-sensitive phenotype. Subsequent analysis showed stable heterozygosity at 16S rRNA position 1408 with a single wild-type allele and a single resistant allele. These results demonstrate that rRNA-mediated mutational resistance to aminoglycosides is recessive.
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Springer, Burkhard, Yishak G. Kidan, Therdsak Prammananan, Kerstin Ellrott, Erik C. Böttger i Peter Sander. "Mechanisms of Streptomycin Resistance: Selection of Mutations in the 16S rRNA Gene Conferring Resistance". Antimicrobial Agents and Chemotherapy 45, nr 10 (1.10.2001): 2877–84. http://dx.doi.org/10.1128/aac.45.10.2877-2884.2001.

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ABSTRACT Chromosomally acquired streptomycin resistance is frequently due to mutations in the gene encoding the ribosomal protein S12,rpsL. The presence of several rRNA operons (rrn) and a single rpsL gene in most bacterial genomes prohibits the isolation of streptomycin-resistant mutants in which resistance is mediated by mutations in the 16S rRNA gene (rrs). Three strains were constructed in this investigation: Mycobacterium smegmatis rrnB,M. smegmatis rpsL 3+, and M. smegmatis rrnB rpsL 3+. M. smegmatis rrnB carries a single functional rrnoperon, i.e., rrnA (comprised of 16S, 23S, and 5S rRNA genes) and a single rpsL +gene; M. smegmatis rpsL 3+ is characterized by the presence of two rrn operons (rrnA and rrnB) and threerpsL + genes; and M. smegmatis rrnB rpsL 3+ carries a single functionalrrn operon (rrnA) and threerpsL + genes. By genetically altering the number of rpsL and rrs alleles in the bacterial genome, mutations in rrs conferring streptomycin resistance could be selected, as revealed by analysis of streptomycin-resistant derivatives of M. smegmatis rrnB rpsL 3+. Besides mutations well known to confer streptomycin resistance, novel streptomycin resistance conferring mutations were isolated. Most of the mutations were found to map to a functional pseudoknot structure within the 530 loop region of the 16S rRNA. One of the mutations observed, i.e., 524G→C, severely distorts the interaction between nucleotides 524G and 507C, a Watson-Crick interaction which has been thought to be essential for ribosome function. The use of the single rRNA allelic M. smegmatis strain should help to elucidate the principles of ribosome-drug interactions.
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Köhler, Gerwald, Wolfgang Ludwig i Karl Heinz Schleifer. "Differentiation of lactococci by rRNA gene restriction analysis". FEMS Microbiology Letters 84, nr 3 (grudzień 1991): 307–12. http://dx.doi.org/10.1111/j.1574-6968.1991.tb04615.x.

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Miyashita, Mika, Takeshi Sakane, Ken-ichiro Suzuki i Yasuyoshi Nakagawa. "16S rRNA gene and 16S-23S rRNA gene internal transcribed spacer sequences analysis of the genus Myxococcus". FEMS Microbiology Letters 282, nr 2 (4.04.2008): 241–45. http://dx.doi.org/10.1111/j.1574-6968.2008.01127.x.

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Paul, Bobby. "Concatenated 16S rRNA sequence analysis improves bacterial taxonomy". F1000Research 11 (19.12.2022): 1530. http://dx.doi.org/10.12688/f1000research.128320.1.

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Background: Microscopic, biochemical, molecular, and computer-based approaches are extensively used to identify and classify bacterial populations. Advances in DNA sequencing and bioinformatics workflows have facilitated sophisticated genome-based methods for microbial taxonomy although sequencing of the 16S rRNA gene is widely employed to identify and classify the bacterial community as a cost-effective and single-gene approach. However, the 16S rRNA sequence-based species identification accuracy is limited by multiple copies of the gene and their higher sequence identity between closely related species. The availability of a large volume of bacterial whole-genome data provided an opportunity to develop comprehensive species-specific 16S rRNA reference libraries. Methods: The 16S rRNA copies were retrieved from the whole genomes in the complete stage at the Genome database. With defined rules, four 16S rRNA gene copy variants were concatenated to develop a species-specific reference library. The sequence similarity search was performed with a web-based BLAST program, and MEGA software was used to construct the phylogenetic tree. Results: Using this approach, species-specific 16S rRNA gene libraries were developed for four closely related Streptococcus species (S. gordonii, S. mitis, S. oralis, and S. pneumoniae). Sequence similarity and phylogenetic analysis using concatenated 16S rRNA copies yielded better resolution than single gene copy approaches. Conclusions: The approach is very effective in classifying genetically related species and may reduce misclassification of bacterial species and genome assemblies.
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Baylis, H. A., i M. J. Bibb. "Transcriptional analysis of the 16S rRNA gene of the rrnD gene set of Streptomyces coelicolor A3(2)". Molecular Microbiology 2, nr 5 (wrzesień 1988): 569–79. http://dx.doi.org/10.1111/j.1365-2958.1988.tb00065.x.

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Rozprawy doktorskie na temat "RRNA gene analysis"

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Lanyon, Clare. "16S rRNA gene sequence analysis of non-methanogenic Archaea in a hypereutrophic freshwater lake". Thesis, University of Liverpool, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.402322.

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Hermansson, Anna. "Ammonia-oxidising bacteria in soil : studies of diversity and abundance using 16S rRNA gene analysis /". Linköping : Univ, 2001. http://www.bibl.liu.se/liupubl/disp/disp2001/tek712s.pdf.

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Wakeman, Jane A. "Human ribosomal RNA : primary structure analysis of the 28S rRNA gene and preliminary studies on the distribution of pseudouridine residues in the 18S and 28S rRNA molecules". Thesis, University of Liverpool, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.317264.

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Calus, Szymon Tomasz. "Evaluation of nanopore-based sequencing technology for gene marker based analysis of complex microbial communities : method development for accurate 16S rRNA gene amplicon sequencing". Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/41086/.

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Nucleic acid sequencing can provide a detailed overview of microbial communities in comparison with standard plate-culture methods. Expansion of high-throughput sequencing (HTS) technologies and reduction in analysis costs has allowed for detailed exploration of various habitats with use of amplicon, metagenomics, and metatranscriptomics approaches. However, due to a capital cost of HTS platforms and requirements for batch analysis, genomics-based studies are still not being used as a standard method for the comprehensive examination of environmental or clinical samples for microbial characterization. This research project investigated the potential of a novel nanopore-based sequencing platform from Oxford Nanopore Technologies (ONT) for rapid and accurate analysis of various environmentally complex samples. ONT is an emerging company that developed the first-ever portable nanopore-based sequencing platform called MinIONTM. Portability and miniaturised size of the device gives an immense opportunity for de-centralised, in-field, and real-time analysis of environmental and clinical samples. Nonetheless, benchmarking of this new technology against the current gold-standard platform (i.e., Illumina sequencers) is necessary to evaluate nanopore data and understand its benefits and limitations. The focus of this study is on the evaluation of nanopore sequencing data: read quality, sequencing errors, alignment quality but also bacterial community structure. For this reason, mock bacterial community samples were generated, sequenced and analysed with use of multiple bioinformatics approaches. Furthermore, this study developed sophisticated library preparation and data analyses methods to enable high-accuracy analysis of amplicon libraries from complex microbial communities for sequencing on the nanopore platform. Besides, the best performing library preparation and data analyses methods were used for analysis of environmental samples and compared to high-quality Illumina metagenomics data. This work opens a new possibility for accurate, in-field amplicon analysis of complex samples with the use of MinIONTM and for the development of autonomous biosensing technology for culture-free detection of pathogenic and non-pathogenic microorganisms in water, soil, food, drinks or blood.
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Al, Masalma Mouhamad. "Molecular and cultural analysis of the bacterial flora associated with brain abscesses". Thesis, Aix-Marseille 2, 2011. http://www.theses.fr/2011AIX20663/document.

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Les abcès cérébraux sont des infections potentiellement mortelles, entraînant souvent des séquelles graves. La prise en charge médicale en reste empirique en raison d’un manque de connaissance approfondie des microorganismes responsables de cette condition. Dans la plupart des laboratoires microbiologiques, le diagnostic d’abcès cérébral est basé sur la culture du pus recueilli chirurgicalement. Malheureusement, cette procédure a de nombreuses limites et ne permet l’identification que d’une petite partie de la population microbienne en cause. L’amplification par PCR et le séquençage du gène codant la fraction 16S de l’ADN ribosomal ont récemment été utilisées pour surmonter les limites de la culture, et ont été démontré leur efficacité dans la documentation des infections bactériennes. Malheureusement, cette procédure présente un degré de discrimination limité en cas d’infection polymicrobienne. Des études métagénomiques de flores complexes de l’homme, basées sur une combinaison de PCR, clonage et séquençage des produits de PCR se sont avérées utiles pour évaluer la diversité bactérienne des flores dentaires, vaginales et intestinales. Nous avons appliqué cette technique à des échantillons d’abcès cérébral pour étudier la flore associée à cette maladie. Dans une première étape, nous avons réalisé une enquête en utilisant la culture et les techniques moléculaires. Le but de cette étude était d’analyser et d’évaluer les bactéries de la flore responsable des abcès cérébraux, en comparant la culture à trois techniques moléculaires basées sur le gène 16S rDNA, incluant le séquençage direct, le clonage suivi de séquençage par méthode de Sanger, et le séquençage direct des produits de PCR par pyroséquençage. Cette enquête a déterminé que la variété des espèces bactériennes associée aux abcès cérébraux est beaucoup plus grande que précédemment décrite, et inclut de nombreuses bactéries anaérobies et des bactéries incultivables de la flore buccale. Cette étude préliminaire a identifié 49 agents bactériens différents, et a permis l’identification de 27 bactéries jamais détectées auparavant dans des abcès du cérébraux, dont 15 n’avaient jamais été cultivées. Un tel nombre d’espèces bactériennes impliquées dans les abcès cérébraux a motivé l’étude de 51 nouveaux spécimens dans le but de décrire plus en détail la flore associée aux abcès cérébraux en fonction de leurs étiologies. Ainsi, nous avons effectué une analyse métagénomique, basé sur le gène 16S rDNA, de 51 patients ayant développé un abcès cérébral. Notre stratégie a été beaucoup plus discriminatoire et a permis à l’identification d’un plus grand nombre de bactéries que la culture et l’amplification et le séquençage direct de l’ANRr 16S. La combinaison des données de 71 patients (20 de la première étude et 51 de la deuxième étude) a permis l’identification de plusieurs associations à l’aide de la méthode de data mining.En outre, notre étude a permis l’identification de deux nouvelles bactéries, la première étant une nouvelle espèce de genre Staphylococcus (Staphylococcus massiliensis) et la seconde étant une bactérie anaérobie qui représente une nouvelle espèce dans un nouveau genre au sein du phylum des Bacteroidetes (Phocaeicola abscesses). En outre, nous avons décrit deux cas inhabituels d’abcès du cerveau, à Mycoplasma hominis après curetage utérin, et à Nocardia carnea chez un greffé rénal. Malgré les limites inhérentes à la procédure de clonage, nos résultats suggèrent que le clonage et le séquençage de gène DNAr 16S est une méthode très performante pour identifier les agents bactériens associés aux abcès cérébraux
Brain abscess is a life-threatening infection with frequent serious sequelae. The medical management remains empirical due to a lack of comprehensive knowledge of the microorganisms responsible for this condition. In most microbiology laboratories the diagnosis of brain abscess is based on culture from pus collected surgically. Unfortunately, this procedure has many limitations and reveals only a small portion of the true microbial population. PCR-amplified 16S rDNA sequencing has recently been used to overcome the limitations of culture-based bacterial detection in brain abscess pus, and it was demonstrated to be effective in the documentation of monomicrobial infections. Unfortunately, this procedure failed to discriminate among polymicrobial floras.Metagenomic studies of complex human floras using a combination of 16S rDNA PCR and cloning-sequencing of PCR products proved useful to evaluate the bacterial diversity of dental, vaginal and intestinal floras. Thus, we applied this technique to brain abscess samples to study the flora associated with this condition. In a first step, we performed an investigation using culture and molecular techniques. The purpose of this investigation was to analyze and evaluate the bacterial flora responsible for brain abscess by comparing standard culture technique to three techniques using 16S rDNA amplification, that is, direct sequencing, multiple sequencing following cloning, and multiple sequencing via high throughput pyrosequencing. This investigation has determined that the variety of brain abscess-associated bacterial species is much larger than previously reported, and it includes many anaerobes and uncultured bacteria from the oral cavity flora. This preliminary study identified 49 distinct brain abscess bacterial agents, and enabled the identification of 27 bacteria never detected before in brain abscess, 15 of which were uncultured.Such a high number of bacterial species involved in brain abscess prompted the study of 51 new specimens in an effort to describe further the flora associated with brain abscesses and their etiologies. Thus, we performed a 16S rDNA-based metagenomic analysis of cerebral abscesses from 51 patients. Our strategy was significantly more discriminatory and enabled the identification of greater number of bacterial taxa, than culture and conventional 16S rDNA PCR/sequencing, respectively. The combination of data from 71 patients (20 from the first study and 51 from the second study) enabled the identification of several associations using the data mining analysis. Also, these studies permitted the identification of two novel bacteria, the first being a novel Staphylococcus species (Staphylococcus massiliensis) and the second being a novel anaerobic bacterium that represents a novel species in a new genus within the phylum Bacteroidetes (Phocaeicola abscesses). In addition, we reported tow unusual cases of brain abscess, the first case was a Mycoplasma hominis brain abscess following uterus curettage and the second case was a Nocardia carnea infection in a kidney transplant recipient patient.Despite limitations inherent to the cloning procedure, our results suggest that cloning and sequencing of PCR-amplified 16S rDNA is a highly valuable method to identify bacterial agents of brain abscesses
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Gustafson, Aubree Marie. "T-RFLP analysis of bacterial 16S rRNA gene sequences isolated from river otter (Lontra canadensis) scat and parasite screening for the presence of Toxoplasma gondii". Scholarly Commons, 2009. https://scholarlycommons.pacific.edu/uop_etds/732.

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In order to analyze the bacterial community of river otter scat (fecal material) at the class level, river otter scat samples were collected from Grizzly Island Wildlife Area (Solano County, CA) and the Cosumnes River Preserve (Sacramento County, CA). DNA was isolated from each sample with the MOBIO PowerSoil™ DNA Isolation Kit and 16S rRNA gene sequences were amplified from each sample. After digestion with Mspl, TRFLPs were analyzed in an ABI Prism™ 310 Genetic Analyzetin triplicate and data peak information from each electropherogram was uploaded into the Phylogenetic Assignment Tool (PAT). Species belonging to the Class Bacilli were the most abundant followed by unclassified species. Two road killed river otters were necropsied to recover brain and blood tissue. DNA was isolated using the Qiagen Tissue DNeasy Kit. Samples from both otters were amplified with a singe tube nested PCR primer set for the detection of the ITS 1 region of Toxoplasma gondii. Scat samples used in the T-RFLP analysis were also tested for the presence ofT. gondii using the same nested primer set. Neither the river otter tissue samples nor any of the scat samples used in this analysis showed evidence of infeGtion with T. gondii.
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Kanso, Sungwan, i n/a. "Molecular Studies of Bacterial Communities in the Great Artesian Basin Aquifers". Griffith University. School of Biomolecular and Biomedical Science, 2004. http://www4.gu.edu.au:8080/adt-root/public/adt-QGU20040219.140509.

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16S rRNA gene analysis has shown that bacterial diversity in the GAB bores studied was limited to the genera Hydrogenobacter in the phylum Aquificae, Thermus in the phylum Deinococcus-Thermus, Desulfotomaculum in the phylum Firmicutes, the alpha-, beta- and gamma-classes of the phylum Proteobacteria and the phylum Nitrospirae. There was no clone closely related to members of the delta-proteobacteria and epsilon-proteobacteria classes detected. The number of bacterial strains directly isolated from the Fairlea and the Cooinda bores were far less than the numbers of distinctive phylotypes detected by the 16S rRNA gene characterisation. In addition none of the bacterial strains directly isolated from the water samples were represented in the 16S rRNA gene clone libraries. Similar discrepancies between the bacterial populations obtained from the 16S rRNA gene analysis and those obtained from direct isolation have been reported in the literature (Dunbar et al., 1999; Kampfer et al., 1996; Suzuki et al., 1997; Ward et al., 1998; Ward et al., 1997). However, in general, the phyla with which the isolates were affiliated were the same as those phyla to which the clones belonged. The environmental changes introduced (by bringing the artesian water up to the surface and exposing it to four types of metal coupons made of carbon steels identified by codes ASTM-A53B, ASTM-A53, AS-1074 and AS-1396 and commonly used in bore casings) led to changes in the bacterial community structures. In general, the species which proliferated in the communities before and after the changes were different. The diversity of the bacterial species in the community decreased following the environmental changes. Clones dominating the clone libraries constructed from newly established bacterial communities also differed from the clones dominating the libraries constructed from the bacterial communities which had existed naturally in the bores. These trends toward change in the bacterial communities were observed at both the Fairlea and the Cooinda bore sites. All four metal types incubated in the Fairlea bore water lost between 3.4 and 4.7% of their original weight. In contrast none of the metals incubated in Cooinda bore water lost weight. Clone library A1 showed that the natural population of the Fairlea bore was dominated by clone A1-3, which represented a novel species related to the isolate boom-7m-04. But after metal incubation (and recording of the metal weight loss), the bacterial community was dominated by clone PKA34B, which has a 95% similarity in its 16S rRNA gene sequence with Desulfotomaculum putei. Desulfotomaculum species are known to cause metal corrosion due to their byproduct H2S. But the low level of phylogenetic relatedness found does not provide enough information to speculate on whether the species represented by clone PKA34B is a member of the genus Desulfotomaculum or not. However, the fact that clone PKA34B dominated the PKA clone library by 50% makes the species it represents a suspected candidate likely to be involved with the metal weight loss at the Fairlea bore. In contrast, clone library 4381 showed that the natural population of the Cooinda bore was dominated by clone 4381-15 representing a species distantly related to a hydrogen oxidiser Hydrogenophaga flava (95% similarity). The dominating clone of the new community formed after metal incubation was clone COO25, which has 99% similarity with Thermus species that have not been reported to be involved with metal corrosion to my knowledge. In this project detection, identification and comparative quantification by 16S rRNA gene-targeted PCR probing with probes 23B and 34B were successfully developed for a Leptothrix-like species and for a Desulfotomaculum-like species represented by clones PKA23B and PKA34B respectively. This method of probing permits a fast, sensitive and reproducible detection, identification and at least a comparative quantification of the bacteria in the environment without the need for culturing. Therefore it is extremely suitable for use in bacterial population monitoring. PCR probing with the 34B probe has a potential commercial use as a means of screening for bores with a potential high risk of corrosion due to this Desulfotomaculum-like species. Direct isolation of bacteria from the GAB water has resulted in the isolation of seven strains from the Fairlea bore and eight from the Cooinda bore. Among these isolates, three novel strains were studied in detail. Reports on the characterisation of strain FaiI4T (T=Type strain) from the Fairlea bore (Kanso & Patel, 2003) and strain CooI3BT from the Cooinda bore have been published (Kanso et al., 2002). The data generated during this project add to our current information and extend our knowledge about the bacterial communities of the GAB's sub-surface environment. This information will provide a basis for further ecological studies of the GAB. Studies on involvement of certain groups of bacteria with the corrosion of metals used in bore casings could provide a foundation for further studies to develop maintenance and managing strategies for the GAB bores.
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Kanso, Sungwan. "Molecular Studies of Bacterial Communities in the Great Artesian Basin Aquifers". Thesis, Griffith University, 2004. http://hdl.handle.net/10072/366613.

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16S rRNA gene analysis has shown that bacterial diversity in the GAB bores studied was limited to the genera Hydrogenobacter in the phylum Aquificae, Thermus in the phylum Deinococcus-Thermus, Desulfotomaculum in the phylum Firmicutes, the alpha-, beta- and gamma-classes of the phylum Proteobacteria and the phylum Nitrospirae. There was no clone closely related to members of the delta-proteobacteria and epsilon-proteobacteria classes detected. The number of bacterial strains directly isolated from the Fairlea and the Cooinda bores were far less than the numbers of distinctive phylotypes detected by the 16S rRNA gene characterisation. In addition none of the bacterial strains directly isolated from the water samples were represented in the 16S rRNA gene clone libraries. Similar discrepancies between the bacterial populations obtained from the 16S rRNA gene analysis and those obtained from direct isolation have been reported in the literature (Dunbar et al., 1999; Kampfer et al., 1996; Suzuki et al., 1997; Ward et al., 1998; Ward et al., 1997). However, in general, the phyla with which the isolates were affiliated were the same as those phyla to which the clones belonged. The environmental changes introduced (by bringing the artesian water up to the surface and exposing it to four types of metal coupons made of carbon steels identified by codes ASTM-A53B, ASTM-A53, AS-1074 and AS-1396 and commonly used in bore casings) led to changes in the bacterial community structures. In general, the species which proliferated in the communities before and after the changes were different. The diversity of the bacterial species in the community decreased following the environmental changes. Clones dominating the clone libraries constructed from newly established bacterial communities also differed from the clones dominating the libraries constructed from the bacterial communities which had existed naturally in the bores. These trends toward change in the bacterial communities were observed at both the Fairlea and the Cooinda bore sites. All four metal types incubated in the Fairlea bore water lost between 3.4 and 4.7% of their original weight. In contrast none of the metals incubated in Cooinda bore water lost weight. Clone library A1 showed that the natural population of the Fairlea bore was dominated by clone A1-3, which represented a novel species related to the isolate boom-7m-04. But after metal incubation (and recording of the metal weight loss), the bacterial community was dominated by clone PKA34B, which has a 95% similarity in its 16S rRNA gene sequence with Desulfotomaculum putei. Desulfotomaculum species are known to cause metal corrosion due to their byproduct H2S. But the low level of phylogenetic relatedness found does not provide enough information to speculate on whether the species represented by clone PKA34B is a member of the genus Desulfotomaculum or not. However, the fact that clone PKA34B dominated the PKA clone library by 50% makes the species it represents a suspected candidate likely to be involved with the metal weight loss at the Fairlea bore. In contrast, clone library 4381 showed that the natural population of the Cooinda bore was dominated by clone 4381-15 representing a species distantly related to a hydrogen oxidiser Hydrogenophaga flava (95% similarity). The dominating clone of the new community formed after metal incubation was clone COO25, which has 99% similarity with Thermus species that have not been reported to be involved with metal corrosion to my knowledge. In this project detection, identification and comparative quantification by 16S rRNA gene-targeted PCR probing with probes 23B and 34B were successfully developed for a Leptothrix-like species and for a Desulfotomaculum-like species represented by clones PKA23B and PKA34B respectively. This method of probing permits a fast, sensitive and reproducible detection, identification and at least a comparative quantification of the bacteria in the environment without the need for culturing. Therefore it is extremely suitable for use in bacterial population monitoring. PCR probing with the 34B probe has a potential commercial use as a means of screening for bores with a potential high risk of corrosion due to this Desulfotomaculum-like species. Direct isolation of bacteria from the GAB water has resulted in the isolation of seven strains from the Fairlea bore and eight from the Cooinda bore. Among these isolates, three novel strains were studied in detail. Reports on the characterisation of strain FaiI4T (T=Type strain) from the Fairlea bore (Kanso & Patel, 2003) and strain CooI3BT from the Cooinda bore have been published (Kanso et al., 2002). The data generated during this project add to our current information and extend our knowledge about the bacterial communities of the GAB's sub-surface environment. This information will provide a basis for further ecological studies of the GAB. Studies on involvement of certain groups of bacteria with the corrosion of metals used in bore casings could provide a foundation for further studies to develop maintenance and managing strategies for the GAB bores.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Biomolecular and Biomedical Sciences
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Kuo, Tai-chih. "The group I ribozyme from the chloroplast rRNA gene of Chlamydomonas reinhardtii : kinetic and structural analysis of the divalent metal requirement and specific interactions with manganese (II) /". Digital version accessible at:, 1998. http://wwwlib.umi.com/cr/utexas/main.

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Harvey, Robert Jr. "Using PCR Amplification and Genetic Sequence Analysis of 18S rRNA Genes to Survey the Microbial Diversity and Distribution of Eukaryotic Microbes Inhabiting Two Thermo-acidic Streams in Yellowstone National Park, Wyoming". ScholarWorks@UNO, 2009. http://scholarworks.uno.edu/td/978.

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A cultivation-independent approach, sequence analysis of 18S rRNA genes PCR-amplified from environmental DNA, was used to explore the diversity and distribution of eukaryotic microbes inhabiting algal mats in two acidic geothermal streams in Yellowstone National Park. The objectives were to: (1) clarify the identity of mat forming algae in Nymph Creek (2) survey microbial species in the Nymph Creek mat over seasonal intervals along a thermal gradient (3) compare microbial species in the Nymph Creek mat with those in Alluvium Creek mats (4) evaluate microbial species in algal mats formed on different substrates in Alluvium Creek. The results show that a novel red alga dominates high temperature regions (~50ºC) of Nymph Creek and two "Chlorella-like" algae predominate the cooler regions (<38ºC). The predominant algae in Alluvium Creek were distinctly different from those in Nymph Creek. Several stramenophiles and fungi were detected in each algal mat.
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Książki na temat "RRNA gene analysis"

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Cheng, Bifang. Cytogenetics of Brassica species and addition lines: Karyotypes, genome analysis and rRNA gene loci. Svalöf, Sweden: Swedish University of Agricultural Sciences, Dept. of Plant Breeding Research, 1996.

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Części książek na temat "RRNA gene analysis"

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Hall, Michael, i Robert G. Beiko. "16S rRNA Gene Analysis with QIIME2". W Methods in Molecular Biology, 113–29. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8728-3_8.

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Comtet-Marre, Sophie, Oshma Chakoory i Pierre Peyret. "Targeted 16S rRNA Gene Capture by Hybridization and Bioinformatic Analysis". W Microbial Environmental Genomics (MEG), 187–208. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2871-3_10.

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Saha, Mousumi, Agniswar Sarkar i Bidyut Bandyopadhyay. "Phylogenetic Characterization of Nitrifying Bacteria Isolated from East Kolkata Wetland". W Proceedings of the Conference BioSangam 2022: Emerging Trends in Biotechnology (BIOSANGAM 2022), 114–22. Dordrecht: Atlantis Press International BV, 2022. http://dx.doi.org/10.2991/978-94-6463-020-6_12.

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AbstractEast Kolkata Wetland (EKW) is an “International Ramsar Site”, famous for broad biodiversity and insightful use of sewage for aquaculture. Native nitrifying bacteria of EKW play a significant role in maintaining water quality and controlling environmental pollution by converting ammonia into nitrate in wastewater. Therefore, the characterization of nitrifying bacteria is important in EKW. Thus, the main focus of this research was to identify and characterize the nitrifying bacteria, investigating their phylogeny and diversity in EKW. 16S rRNA and functional genes analysis may help in the proper evaluation of composition and distribution of nitrifying bacteria in some water bodies in EKW, which has not yet been explored. Molecular and phylogenetic characterization was targeted and achieved through 16S rRNA and functional gene analysis, followed by computational estimation. Resulted sequences were analysed to gain insight into the knowledge for global and local taxonomic orientation. Hence, a model can be created for characterizing the dynamics of nitrifying bacteria in wastewater treatment and sustainable aquaculture in different water bodies of EKW. Graphical Abstract
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Merkl, Philipp E., Christopher Schächner, Michael Pilsl, Katrin Schwank, Kristin Hergert, Gernot Längst, Philipp Milkereit, Joachim Griesenbeck i Herbert Tschochner. "Analysis of Yeast RNAP I Transcription of Nucleosomal Templates In Vitro". W Ribosome Biogenesis, 39–59. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2501-9_3.

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AbstractNuclear eukaryotic RNA polymerases (RNAPs) transcribe a chromatin template in vivo. Since the basic unit of chromatin, the nucleosome, renders the DNA largely inaccessible, RNAPs have to overcome the nucleosomal barrier for efficient RNA synthesis. Gaining mechanistical insights in the transcription of chromatin templates will be essential to understand the complex process of eukaryotic gene expression. In this article we describe the use of defined in vitro transcription systems for comparative analysis of highly purified RNAPs I–III from S. cerevisiae (hereafter called yeast) transcribing in vitro reconstituted nucleosomal templates. We also provide a protocol to study promoter-dependent RNAP I transcription of purified native 35S ribosomal RNA (rRNA) gene chromatin.
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Duduk, Bojan, Samanta Paltrinieri, Ing-Ming Lee i Assunta Bertaccini. "Nested PCR and RFLP Analysis Based on the 16S rRNA Gene". W Methods in Molecular Biology, 159–71. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-089-2_14.

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Barrios Hernández, Mary Luz. "Eukaryotic community characterisation by 18s rRNA gene analysis in full-scale systems". W Pathogen removal in aerobic granular sludge treatment systems, 91–112. London: CRC Press, 2022. http://dx.doi.org/10.1201/9781003231622-5.

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Lawley, Blair, i Gerald W. Tannock. "Analysis of 16S rRNA Gene Amplicon Sequences Using the QIIME Software Package". W Methods in Molecular Biology, 153–63. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-6685-1_9.

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Stackebrandt, Erko. "Pitfalls of PCR-Based rRNA Gene Sequence Analysis: An Update on Some Parameters". W Handbook of Molecular Microbial Ecology I, 129–33. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2011. http://dx.doi.org/10.1002/9781118010518.ch16.

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Martini, Marta, Kristi D. Bottner-Parker i Ing-Ming Lee. "PCR-Based Sequence Analysis on Multiple Genes Other than 16S rRNA Gene for Differentiation of Phytoplasmas". W Phytoplasmas, 97–115. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8837-2_8.

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Mohannath, Gireesha, i Craig S. Pikaard. "Analysis of rRNA Gene Methylation in Arabidopsis thaliana by CHEF-Conventional 2D Gel Electrophoresis". W The Nucleolus, 183–202. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3792-9_14.

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Streszczenia konferencji na temat "RRNA gene analysis"

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Silva, M. L. R. Bastos, M. C. C. Pereira Lyra, J. Paula Oliveira, H. Almeida Burity i G. M. Campos-Takaki. "Microbial diversity in Chromobacterium violaceum determined by 16S rRNA gene analysis". W Proceedings of the II International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld2007). WORLD SCIENTIFIC, 2009. http://dx.doi.org/10.1142/9789812837554_0063.

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Aleev, Vladislav S., i Elena G. Plotnikova. "GENETIC AND ECOLOGICAL-PHYSIOLOGICAL CHARACTERISTICS OF BACTERIA OF THE GENUS SALINISPHAERA (CLASS GAMMAPROTEOBACTERIA) ISOLATED FROM THE VERKHNEKAMSKY SALT MINER". W Фундаментальные и прикладные исследования в биологии и экологии. Пермский государственный национальный исследовательский университет, 2021. http://dx.doi.org/10.17072/fpibe-2021-4-7.

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5 strains of halophilic bacteria were isolated from clay deposits of brine-diverting workings and brine tanks of the mine of the Verkhnekamskoye salt deposit (Solikamsk, Perm Territory). Because of phylogenetic analysis based on a comparison of the 16S rRNA gene sequences, it was found that the isolated cultures are members of the Salinisphaeraceae family. Three halophilic strains SHV2, RV14, and SWV1 had a similarity with the closest type strain of the Salinisphaera hydrothermalis species at the level of 95.94-96.62% (16S rRNA gene), which indicates that these strains belong to a new taxon. All isolated bacteria are halophiles: they grow at high salinity (up to 270-300 g / l NaCl).
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Deljou, A., K. Mousavi, A. Ghasemi i H. Rahimian. "Identification of Mycobacterium sp. as Alfalfa endophytes using 16s rRNA gene sequence analysis". W Proceedings of the II International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld2007). WORLD SCIENTIFIC, 2009. http://dx.doi.org/10.1142/9789812837554_0013.

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Yamasaki, Kei, Kazuhiro Yatera, Toshinori Kawanami, Kazumasa Fukuda, Shingo Noguchi, Shuya Nagata, Chinatsu Nishida i in. "The Bacteriological Incidence Of Community-Acquired Pneumonia Using Clone Analysis Of 16S RRNA Gene". W American Thoracic Society 2012 International Conference, May 18-23, 2012 • San Francisco, California. American Thoracic Society, 2012. http://dx.doi.org/10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a5243.

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Kawanami, Toshinori, Shingo Noguchi, Kentarou Akata, Keisuke Naito, Tsutomu Takaki, Keishi Oda, Minako Hanaka, Takashi Kido, Hiroshi Mukae i Kazuhiro Yatera. "Risk factors associated with “PES pathogens” in community-onset pneumonia: 16S rRNA gene analysis of BALF". W ERS International Congress 2017 abstracts. European Respiratory Society, 2017. http://dx.doi.org/10.1183/1393003.congress-2017.oa471.

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Naito, Keisuke, Shingo Noguchi, Kazuhiro Yatera, Kentarou Akata, Chinatsu Nishida, Kei Yamasaki, Takeshi Orihashi i in. "The bacteriological incidence of lung abscess using clone library analysis of 16S rRNA gene in bronchoalveolar lavage". W Annual Congress 2015. European Respiratory Society, 2015. http://dx.doi.org/10.1183/13993003.congress-2015.pa5040.

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Eltai, Nahla O., Sara H. Al-Hadidi, Asmaa A. Al Than, Sanjay H. Doiphode i Hadi M. Yassine. "Salmonellosis among Pediatric Population in Qatar: Prevalence, Antibiotic Resistance and Molecular Epidemiology". W Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0126.

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Objectives: This study aims to characterize at the molecular level the genes encoding resistance in Salmonella and explain the molecular mechanisms underlying resistance to ceftriaxone, cefepime, amoxicillin-clavulanate, tetracycline, trimethoprim-sulfamethoxazole, chloramphenicol, colistin and azithromycin in Salmonella. It aims as well to characterize the 16S rRNA gene region by restriction fragment length polymorphism (RFLP) to investigate if this region constitutes an appropriate ‘coincidental’ marker to distinguish important pathogenic Salmonella species. Finally, determine the lineages of Salmonella species and evolutionary relationships among bacteria classified within the same genus. Methodology: 246 Salmonella isolates were collected from children under 16 years old during Jan. 2018 - Dec 2019, presented with gastroenteritis at Hamad Medical Corporation. Isolates were tested for antibiotic susceptibility against nineteen relevant antibiotics using E-test. Isolates that harbor antibiotic resistance were confirmed using PCR specific primers for 38 genes. In addition, the variable region of class 1 and 2 integrons were identified by PCR among amoxicillin-clavulanate (AMC) resistant samples. RFLP targeting16S rRNAwas performed using seven restriction enzymes including AluI, Bgl I, Bgl II, EcoR I, SmaI, Hinf I & Hae III. Results: Resistance was detected against 15 antibiotics and (38.2%) of isolates were resistant to at least one antibiotic. Overall, high resistance was reported to tetracycline (23.9%), ampicillin (21.1%), AMC (18.7%) and sulfamethoxazoletrimethoprim (13%). Further, 22.4% of the isolates were multidrug-resistant (MDR), with 4.1% being ESBL producers. 90 % of ESBL producers had one of bla CTX-M-Group. Class (1) AMC resistant samples showed the highest resistance to different antibiotics. 16S rRNA-RFLP analysis divided Salmonella isolates into two main groups. Conclusion: Our results indicate a high antimicrobial resistance pattern of Salmonella, which necessities the development of regulatory programs to combats antimicrobial resistance. In particular, our results showed high resistance to Class (1) AMC cassette that involves the transmission and expression of the resistance. This might lead to a concern of increased multidrug resistance in the future. This study provides evidence guidance to activate and implement the pillars of an antimicrobial stewardship program in animal and human health to reduce MDR salmonellosis.
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Kawanami, Toshinori, Shingo Noguchi, Kei Yamasaki, Ryosuke Hata, Hiroaki Ikegami, Kaori Kato, Hiroshi Mukae i Kazuhiro Yatera. "Relationship between a detection of anaerobes in the lung and poor oral hygiene; using the bacterial floral analysis of 16S rRNA gene". W ERS International Congress 2019 abstracts. European Respiratory Society, 2019. http://dx.doi.org/10.1183/13993003.congress-2019.oa5145.

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Guro, P., V. Safronova, A. Sazanova, I. Kuznetsova, A. Belimov, V. Yakubov, E. Chirak, A. Afonin, E. Andronov i I. Tikhonovich. "Rhizobial microsymbionts of the narrowly endemic Oxytropis species growing in Kamchatka possess a set of genes that are associated with T3SS and T6SS secretion systems and can affect the development of symbiosis". W 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.099.

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A collection of rhizobial strains isolated from root nodules of the narrowly endemic legume species Oxytropis erecta, O. anadyrensis, O. kamtschatica and O. pumilio growing on the Kamchatka Peninsula (Russian Federation) was obtained. Analysis of the 16S rRNA gene sequence showed a significant diversity of isolates belonging to the families Rhizobiaceae (Rhizobium), Phyllobacteriaceae (Mesorhizobium, Phyllobacterium) and Bradyrhizobiaceae (Bosea, Tardiphaga). Pairs of taxonomically different strains in various combinations were isolated from some nodules of Oxytropis plants. Plant nodulation assays showed that only strains belonging to the genus Mesorhizobium (M. jarvisii, M. loti and M. huakuii) could form nitrogen-fixing nodules. The nitrogen-fixing activity of the strains was more associated with the host plant than with the species of strains. The whole genome sequences analysis showed that the strains M. loti 582 and M. huakuii 583 possessed symbiotic genes necessary for the formation of effective symbiosis and grouped into Sym-clusters. In contrast, the strain T. robiniae 581 had only a reduced number of fix genes, while the strains Phyllobacterium sp. 628 and R. lusitanum 629 possesed only individual symbiotic genes, which obviously did not participate in the formation of nodules. It was also stated that the strains M. loti 582 and M. huakuii 583 had a significantly larger set of genes related to the secretion systems T3SS and T6SS that can affect the host specificity of strains, compared with 6 commercial strains used as reference. These two strains formed nodules of two types (typical elongated and atypical rounded) on Oxytropis plants. We suggest that a possible cause of the observed phenomenon is the availability of different nodulation strategies in these strains (dependent and independent of Nod-factors). Thus, as a result of studying the collection of strains isolated from the narrow endemic species of Kamchatka Oxytropis, interesting objects were selected to study the functions of the T3SS and T6SS genes, and their role in the development of rhizobia-legume symbiosis. The prospects of using strains with gene systems for both symbiotic and non-symbiotic nodulation to enhance the efficiency of plant-microbe interactions by expanding the host specificity and increasing the efficiency of nodulation are discussed.
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Hidayat, Habibi. "Analysis of 16S rRNA gene lactic acid bacteria (LAB) isolate from Markisa fruit (Passiflora sp.) as a producer of protease enzyme and probiotics". W PROCEEDINGS FROM THE 14TH INTERNATIONAL SYMPOSIUM ON THERAPEUTIC ULTRASOUND. Author(s), 2017. http://dx.doi.org/10.1063/1.4978183.

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Raporty organizacyjne na temat "RRNA gene analysis"

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Zchori-Fein, Einat, Judith K. Brown i Nurit Katzir. Biocomplexity and Selective modulation of whitefly symbiotic composition. United States Department of Agriculture, czerwiec 2006. http://dx.doi.org/10.32747/2006.7591733.bard.

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Whiteflies are sap-sucking insects that harbor obligatory symbiotic bacteria to fulfill their dietary needs, as well as a facultative microbial community with diverse bacterial species. The sweetpotato whitefly Bemisia tabaci (Gennadius) is a severe agricultural pest in many parts of the world. This speciesconsists of several biotypes that have been distinguished largely on the basis of biochemical or molecular diagnostics, but whose biological significance is still unclear. The original objectives of the project were (i) to identify the specific complement of prokaryotic endosymbionts associated with select, well-studied, biologically and phylogeographically representative biotypes of B. tabaci, and (ii) to attempt to 'cure’ select biotypes of certain symbionts to permit assessment of the affect of curing on whitefly fitness, gene flow, host plant preference, and virus transmission competency.To identify the diversity of bacterial community associated with a suite of phylogeographically-diverseB. tabaci, a total of 107 populations were screened using general Bacteria primers for the 16S rRNA encoding gene in a PCR. Sequence comparisons with the available databases revealed the presence of bacteria classified in the: Proteobacteria (66%), Firmicutes (25.70%), Actinobacteria (3.7%), Chlamydiae (2.75%) and Bacteroidetes (<1%). Among previously identified bacteria, such as the primary symbiont Portiera aleyrodidarum, and the secondary symbionts Hamiltonella, Cardinium and Wolbachia, a Rickettsia sp. was detected for the first time in this insect family. The distribution, transmission, and localization of the Rickettsia were studied using PCR and fluorescence in situ hybridization (FISH). Rickettsia was found in all 20 Israeli B. tabaci populations screened as well as some populations screened in the Arizona laboratory, but not in all individuals within each population. FISH analysis of B. tabaci eggs, nymphs and adults, revealed a unique concentration of Rickettsia around the gut and follicle cells as well as its random distribution in the haemolymph, but absence from the primary symbiont housing cells, the bacteriocytes. Rickettsia vertical transmission on the one hand and its partial within-population infection on the other suggest a phenotype that is advantageous under certain conditions but may be deleterious enough to prevent fixation under others.To test for the possible involvement of Wolbachia and Cardiniumin the reproductive isolation of different B. tabacibiotypes, reciprocal crosses were preformed among populations of the Cardinium-infected, Wolbachia-infected and uninfected populations. The crosses results demonstrated that phylogeographically divergent B. tabaci are reproductively competent and that cytoplasmic incompatibility inducer-bacteria (Wolbachia and Cardinium) both interfered with, and/or rescued CI induced by one another, effectively facilitating bidirectional female offspring production in the latter scenario.This knowledge has implications to multitrophic interactions, gene flow, speciation, fitness, natural enemy interactions, and possibly, host preference and virus transmission. Although extensive and creative attempts undertaken in both laboratories to cure whiteflies of non-primary symbionts have failed, our finding of naturally uninfected individuals have permitted the establishment of Rickettsia-, Wolbachia- and Cardinium-freeB. tabaci lines, which are been employed to address various biological questions, including determining the role of these bacteria in whitefly host biology.
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Ostersetzer-Biran, Oren, i Jeffrey Mower. Novel strategies to induce male sterility and restore fertility in Brassicaceae crops. United States Department of Agriculture, styczeń 2016. http://dx.doi.org/10.32747/2016.7604267.bard.

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Abstract Mitochondria are the site of respiration and numerous other metabolic processes required for plant growth and development. Increased demands for metabolic energy are observed during different stages in the plants life cycle, but are particularly ample during germination and reproductive organ development. These activities are dependent upon the tight regulation of the expression and accumulation of various organellar proteins. Plant mitochondria contain their own genomes (mtDNA), which encode for rRNAs, tRNAs and some mitochondrial proteins. Although all mitochondria have probably evolved from a common alpha-proteobacterial ancestor, notable genomic reorganizations have occurred in the mtDNAs of different eukaryotic lineages. Plant mtDNAs are notably larger and more variable in size (ranging from 70~11,000 kbp in size) than the mrDNAs in higher animals (16~19 kbp). Another unique feature of plant mitochondria includes the presence of both circular and linear DNA fragments, which undergo intra- and intermolecular recombination. DNA-seq data indicate that such recombination events result with diverged mitochondrial genome configurations, even within a single plant species. One common plant phenotype that emerges as a consequence of altered mtDNA configuration is cytoplasmic male sterility CMS (i.e. reduced production of functional pollen). The maternally-inherited male sterility phenotype is highly valuable agriculturally. CMS forces the production of F1 hybrids, particularly in predominantly self-pollinating crops, resulting in enhanced crop growth and productivity through heterosis (i.e. hybrid vigor or outbreeding enhancement). CMS lines have been implemented in some cereal and vegetables, but most crops still lack a CMS system. This work focuses on the analysis of the molecular basis of CMS. We also aim to induce nuclear or organellar induced male-sterility in plants, and to develop a novel approach for fertility restoration. Our work focuses on Brassicaceae, a large family of flowering plants that includes Arabidopsis thaliana, a key model organism in plant sciences, as well as many crops of major economic importance (e.g., broccoli, cauliflower, cabbage, and various seeds for oil production). In spite of the genomic rearrangements in the mtDNAs of plants, the number of genes and the coding sequences are conserved among different mtDNAs in angiosperms (i.e. ~60 genes encoding different tRNAs, rRNAs, ribosomal proteins and subunits of the respiratory system). Yet, in addition to the known genes, plant mtDNAs also harbor numerous ORFs, most of which are not conserved among species and are currently of unknown function. Remarkably, and relevant to our study, CMS in plants is primarily associated with the expression of novel chimericORFs, which likely derive from recombination events within the mtDNAs. Whereas the CMS loci are localized to the mtDNAs, the factors that restore fertility (Rfs) are identified as nuclear-encoded RNA-binding proteins. Interestingly, nearly all of the Rf’s are identified as pentatricopeptide repeat (PPR) proteins, a large family of modular RNA-binding proteins that mediate several aspects of gene expression primarily in plant organelles. In this project we proposed to develop a system to test the ability of mtORFs in plants, which are closely related to known CMS factors. We will induce male fertility in various species of Brassicaceae, and test whether a down-relation in the expression of the recombinantCMS-genes restores fertility, using synthetically designed PPR proteins.
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