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Registros recuperados: 28
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A cost-effective melting temperature assay for the detection of single-nucleotide polymorphism in the MBL2 gene of HIV-1-infected children BJMBR
Arraes,L.C.; de Souza,P.R.; Bruneska,D.; Castelo Filho,A.; de Souza Cavada,B.; de Lima Filho,J.L.; Crovella,S..
We report a fast (less than 3 h) and cost-effective melting temperature assay method for the detection of single-nucleotide polymorphisms in the MBL2 gene. The protocol, which is based on the Corbett Rotor Gene real time PCR platform and SYBR Green I chemistry, yielded, in the cohorts studied, sensitive (100%) and specific (100%) PCR amplification without the use of costly fluorophore-labeled probes or post-PCR manipulation. At the end of the PCR, the dissociation protocol included a slow heating from 60º to 95ºC in 0.2ºC steps, with an 8-s interval between steps. Melting curve profiles were obtained using the dissociation software of the Rotor Gene-3000 apparatus. Samples were analyzed in duplicate and in different PCR runs to test the reproducibility of...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Melting temperature assay; Real time PCR; MBL2; Polymorphisms; HIV-1-positive children.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000600003
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A relationship between antimicrobial peptide gene expression and capacity of a selected shrimp line to survive a Vibrio infection ArchiMer
De Lorgeril, Julien; Gueguen, Yannick; Goarant, Cyrille; Goyard, Emmanuel; Mugnier, Chantal; Fievet, Julie; Piquemal, D; Bachere, Evelyne.
Understanding of antimicrobial defence mechanisms of penaeid shrimp should help in the design of efficient strategies for the management and disease control in aquaculture. In this study, we have specifically analysed the expression in circulating hemocytes of antimicrobial peptides (AMPs) encoding genes, such as PEN2 and PEN3, ALF, crustin, lysozyme and a putative cysteine-rich peptide. We evidenced a relationship between the level of expression of some AMPs and the successful response of the shrimp, Litopenaeus stylirostris, to circumvent a pathogenic Vibrio penaeicida infection. Additionally, significant differences in some AMP transcript amounts are evidenced between control, non-selected shrimp line and the third generation breeding of shrimp selected...
Tipo: Text Palavras-chave: Real time PCR; Cysteine rich peptide; Crustin; Anti LPS factor; Lysozyme; Penaeidins; Immune response; Vibrio penaeicida; Penaeid; Decapoda; Crustacean.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-4524.pdf
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Analysis of gene expression in Kalanchoe daigremontiana leaves during plantlet formation under drought stress Electron. J. Biotechnol.
Zhong,Tianxiu; Zhu,Chen; Zeng,Huiming; Han,Liebao.
Background: Kalanchoe daigremontiana is an attractive model system for the study of the molecular mechanisms of somatic embryogenesis and organogenesis competence due to its formation of plantlets with adventitious roots on the leaf margins that are derived from somatic embryos. The suppression subtractive hybridization technique was used to investigate gene expression during asexual reproduction. Leaves from plants subjected to drought stress provided the source of ‘Tester’ DNA, and leaves from plants grown under normal conditions provided the ‘Driver’ DNA for subtractive hybridization. Results: A total of 481 high quality ESTs were generated, which clustered into 390 unigenes. Of these unigenes, 132 grouped into 12...
Tipo: Journal article Palavras-chave: Bioinformatics analysis; Real time PCR; Suppression subtractive hybridization.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600004
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Comparative Analysis of C-repeat Binding Factors (CBFs) in Tomato and Arabidopsis BABT
Heidari,Parviz.
Abstract Cold stress is one of the limiting factors of plant production that plants use different mechanisms for cold tolerance. CBF genes play critical role to regulate the cold responsive genes. To better understand of CBF gene functions, the tomato-CBFs and Arabidopsis-CBFs were evaluated using bioinformatics tools, and finally the expression patterns of SlCBF1 gene were analyzed under 10 and 4˚C in two contrasting tomato species (Solanum lycopersicum and S. habrochaites). The different cis regulatory elements were observed in promoter region of SlCBF1 and AtCBF1 genes, and ICE1, COR and HOS1 proteins exhibited high interaction with CBFs. The results of Real time PCR of SlCBF1 exhibited that under 10 and 4 ˚C, SlCBF1 was down regulated in cold sensitive...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cold stress; CBF genes; Real time PCR; Tomato; Bioinformatics analysis.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100208
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Comparison of nine DNA extraction methods for the diagnosis of bovine tuberculosis by real time PCR Ciência Rural
Moura,André; Hodon,Mikael Arrais; Soares Filho,Paulo Martins; Issa,Marina de Azevedo; Oliveira,Ana Paula Ferreira de; Fonseca Júnior,Antônio Augusto.
ABSTRACT: Bovine tuberculosis is an infectious disease with a high impact on the cattle industry, particularly in developing countries. PCR is a very sensitive method for detection of infectious agents, but the sensitivity of molecular diagnosis is largely dependent on the efficiency of the DNA extraction methods. The objective of this study was to evaluate DNA extraction methods for direct detection of Mycobacterium bovis in bovine tissue. Nine commercial kits for DNA extraction were evaluated when combined with two real time PCRs. The DNeasy Blood & Tissue Kit from QIAGEN showed better performance and sensitivity followed by the DNA Mini Kit RBC and FTA Elute Micro Card. Results suggested that, even when the analytical sensitivity of the qPCR is...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine tuberculosis; DNA extraction; Real time PCR.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000701223
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Detección de Erwinia chrysanthemi pv zeae (Sabet 1954) Victoria et al., 1975 en maíz en Morelos, México. Colegio de Postgraduados
Durán Peralta, Elisa.
La “pudrición del tallo del maíz” causada por Erwinia chrysanthemi pv zeae es una de las principales enfermedades del maíz (Zea mays L.) en los países tropicales y subtropicales. En el presente trabajo se realizó la extracción de ADN del patógeno y se realizó PCR utilizando los iniciadores ADE1 y ADE2, diseñados para E. chrysanthemi (Burkh.) Young et al. 1978, que amplifican un fragmento de 420pb del gen pelADE. Se demostró que estos también pueden ser utilizados para su detección por PCR en tiempo real con resultados similares a los citados en la literatura; sin embargo no pueden ser empleados para la cuantificación de la bacteria debido a la formación de estructuras secundarias. Mediante la caracterización fisiológica, bioquímica y análisis...
Palavras-chave: Zea mays L.; Erwinia chrysanthemi pv zeae; Pudrición del tallo del maíz; PCR en tiempo real; Gen pe/ADE; Corn stalk rot; Real time PCR; Gene Pe/ADE; Fitopatología; Maestría.
Ano: 2011 URL: http://hdl.handle.net/10521/555
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Detección de Erwinia chrysanthemi pv zeae (Sabet 1954) Victoria et al., 1975 en maíz en Morelos, México. Colegio de Postgraduados
Durán Peralta, Elisa.
La “pudrición del tallo del maíz” causada por Erwinia chrysanthemi pv zeae es una de las principales enfermedades del maíz (Zea mays L.) en los países tropicales y subtropicales. En el presente trabajo se realizó la extracción de ADN del patógeno y se realizó PCR utilizando los iniciadores ADE1 y ADE2, diseñados para E. chrysanthemi (Burkh.) Young et al. 1978, que amplifican un fragmento de 420pb del gen pelADE. Se demostró que estos también pueden ser utilizados para su detección por PCR en tiempo real con resultados similares a los citados en la literatura; sin embargo no pueden ser empleados para la cuantificación de la bacteria debido a la formación de estructuras secundarias. Mediante la caracterización fisiológica, bioquímica y análisis...
Palavras-chave: Zea mays L.; Erwinia chrysanthemi pv zeae; Pudrición del tallo del maíz; PCR en tiempo real; Gen pe/ADE; Corn stalk rot; Real time PCR; Gene Pe/ADE; Fitopatología; Maestría.
Ano: 2011 URL: http://hdl.handle.net/10521/555
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Different methods of real-time PCR for detection of pseudorabies virus Ciência Rural
Nonaka,Carolina Kymie Vasques; Fonseca Junior,Antônio Augusto; Guedes,Estefânia Oliveira; D´Ambros,Régia Maria; Lima,Graciela Kunrath; Camargos,Marcelo Fernandes; Heinemann,Marcos Bryan.
ABSTRACT: Pseudorabies (PR) is a highly contagious viral disease of great animal health and economic importance in swine industry. The aim of this study was to evaluate different genomic regions, real-time PCR chemistries and equipment for the molecular diagnosis of PR. Eight primer pairs targeting four genes (gB, gC, gE, gD), three different qPCR chemistries (SybrGreen, hydrolysis probes and plexor) and two equipment (ABI7500, Rotorgene 3000) were evaluated. Oligonucleotides targeting gB using hydrolysis probes showed the best performance after evaluating efficiency (99%), the detection limit (10-1.5 TCID50 mL-1) and diagnostic sensitivity and; therefore, those primers were selected for performance verification factors such as repeatability,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Real time PCR; Pseudorabies; Diagnose.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782017000300452
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Differential gene expression and mitotic cell analysis of the drought tolerant soybean (Glycine max L. Merrill Fabales, Fabaceae) cultivar MG/BR46 (Conquista) under two water deficit induction systems Genet. Mol. Biol.
Martins,Polyana K.; Jordão,Berenice Q.; Yamanaka,Naoki; Farias,José R.B.; Beneventi,Magda A.; Binneck,Eliseu; Fuganti,Renata; Stolf,Renata; Nepomuceno,Alexandre L..
Drought cause serious yield losses in soybean (Glycine max), roots being the first plant organ to detect the water-stress signals triggering defense mechanisms. We used two drought induction systems to identify genes differentially expressed in the roots of the drought-tolerant soybean cultivar MG/BR46 (Conquista) and characterize their expression levels during water deficit. Soybean plants grown in nutrient solution hydroponically and in sand-pots were submitted to water stress and gene expression analysis was conducted using the differential display (DD) and real time polymerase chain reaction (PCR) techniques. Three differentially expressed mRNA transcripts showed homology to the Antirrhinum majus basic helix-loop-helix transcription factor bHLH, the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell division; Differential display; Gene expression; Real time PCR; Soybean; Water stress.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000300019
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Early remodeling of rat cardiac muscle induced by swimming training BJMBR
Verzola,R.M.M.; Mesquita,R.A.; Peviani,S.; Ramos,O.H.P.; Moriscot,A.S.; Perez,S.E.A.; Selistre-de-Araújo,H.S..
The aim of the present investigation was to study the effect of acute swimming training with an anaerobic component on matrix metallopeptidase (MMP) activity and myosin heavy chain gene expression in the rat myocardium. Animals (male Wistar rats, weighing approximately 180 g) were trained for 6 h/day in 3 sessions of 2 h each for 1 to 5 consecutive days (N = 5 rats per group). Rats swam in basins 47 cm in diameter and 60 cm deep filled with water at 33 to 35ºC. After the training period a significant increase (P < 0.05) was observed in the heart weight normalized to body weight by about 22 and 35% in the groups that trained for 96 and 120 h, respectively. Blood lactate levels were significantly increased (P < 0.05) in all groups after all training...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Myosin heavy chain expression; Training; Real time PCR; Rat myocardium; Muscle hypertrophy.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000500009
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Etude des effets de polluants sur les paramètres hémocytaires de l'huître creuse, Crassostrea gigas - Interactions entre environnement, mécanismes de défense et maladies infectieuses ArchiMer
Gagnaire, Beatrice.
Shellfish industry is mostly realized in estuary zones, which are subjected to pollutions due to anthropic activities. The harmful effects of pollutants on animals inhabiting these estuarine zones are poorly known. Among these animals, Pacific oyster, Crassostrea gigas, may represent a model because they are sedentary and they filter water intensively. Among all physiological functions possibly disturbed by pollutants, defence mechanisms are poorly studied in bivalves. Moreover, animals presenting impaired defence mechanisms may be more sensitive to infectious diseases. In this context, effects of pollutants on hemocyte functions and on sensitiveness to diseases were tested in C. gigas. After adjusting the protocols for the monitoring of hemocyte...
Tipo: Text Palavras-chave: Real time PCR; Flow cytometry; Pesticides; Pollutants; Phagocytosis; Hemocytes; Crassostrea gigas; Pacific oyster; Pathology; Immunotoxicology; PCR en temps réel; Cytométrie de flux; Pesticides; Polluants; Phagocytose; Hémocytes; Crassostrea gigas; Huître creuse; Pathologie; Immunotoxicologie.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/these-730.pdf
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Expression analysis for genes involved in arachidonic acid biosynthesis in Mortierella alpina CBS 754.68 BJM
Samadlouie,Hamid-Reza; Hamidi-Esfahani,Zohreh; Alavi,Seyed-Mehdi; Varastegani,Boshra.
The time courses for production of fungal biomass, lipid, phenolic and arachidonic acid (ARA) as well as expression of the genes involved in biosynthesis of ARA and lipid were examined in Mortierella alpina CBS 754.68. A significant increase in the arachidonic acid content in lipids that coincided with reduced levels of lipid was obtained. Reduced gene expression occurred presumably due to the steady reduction of carbon and nitrogen resources. However, these energy resources were inefficiently compensated by the breakdown of the accumulated lipids that in turn, induced up-regulated expression of the candidate genes. The results further indicated that the expression of the GLELO encoding gene is a rate-limiting step in the biosynthesis of ARA in the early...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mortierella alpina; Arachidonic acid; Lipid; Gene expression; Real time PCR.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200010
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First description of French V. tubiashii strains pathogenic to mollusk: I. Characterization of isolates and detection during mortality events ArchiMer
Travers, Marie-agnes; Mersni Achour, Rachida; Haffner, Philippe; Tourbiez, Delphine; Cassone, Anne-laure; Morga, Benjamin; Doghri, Ibtissem; Garcia, Celine; Renault, Tristan; Fruitier-arnaudin, Ingrid; Saulnier, Denis.
Nine dominant bacterial isolates were obtained from different batches of Crassostrea gigas spat experiencing high mortality rates in a French experimental hatchery/nursery in 2007. Using phenotypic analysis combined with multilocus sequence analysis, the isolates were shown to be genetically close to the Vibrio tubiashii type strain. Based on (1) analyses of the recA gene sequences; (2) the results of DNA–DNA hybridization assays between 07/118 T2 (LMG 27884 = CECT 8426), which is a representative strain, and the V. tubiashii type strain (69%); and (3) phenotypic traits, the bacteria were classified in a group close to American V. tubiashii strain. Its virulence (70% of mortalities) and the toxicity of the extracellular products of 07/118 T2 was...
Tipo: Text Palavras-chave: Crassostrea gigas Pathogenicity Extracellular products Polyphasic approach; Real time PCR.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00189/30028/28743.pdf
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Identification of a splicing coactivator gene that affects the production of ochratoxin a in Aspergillus carbonarius BABT
Lunardi,Lígia Uno; Guembarovski,Roberta Losi; Hanai,Luiz Ricardo; Cristiano,Valderi; Vieira,Maria Lucia Carneiro; Sartori,Daniele; Fungaro,Maria Helena Pelegrinelli.
Ochratoxin A is a mycotoxin produced by some fungi species. Among them, Aspergillus carbonarius is considered a powerful producer. Genes involved in the ochratoxin A biosynthesis pathway have been identified in some producer species. However, there are few studies that purpose to identify these genes in A. carbonarius. The use of insertion mutants to identify genes associated with certain properties has been increased in the literature. In this work, the region of T-DNA integration was investigated in one A. carbonarius ochratoxin-defective mutant previously obtained by Agrobacterium tumefaciens-mediated transformation, in order to find an association between interrupted gene and the biosynthesis of ochratoxin A. The integration occurred in a gene that...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Ochratoxigenic fungi; Mycotoxin; Real time PCR; Splicing coactivator protein.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700018
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Identification of genes from flat oyster Ostrea edulis as suitable housekeeping genes for quantitative real time PCR ArchiMer
Morga, Benjamin; Arzul, Isabelle; Faury, Nicole; Renault, Tristan.
Bonamia ostreae is an intrahaemocytic protozoan affecting Ostrea edulis. The parasite multiplies within haemocytes without being degraded and involves changes in cellular activities. Studies aiming at better understanding host response to a pathogen at the transcriptome levels are frequently based on the use of real time PCR assays, which require some reference genes. However, very few sequence data is available for O. edulis in public databases. Subtracted cDNA libraries were constructed from the O. edulis haemocytes in order to identify genes involved in host reactions against the parasite and quantitative real time PCR assays were developed to study expression of these genes. In this context, identification of reference genes and study of their...
Tipo: Text Palavras-chave: Real time PCR; Housekeeping genes; Haemocytes; Ostrea edulis; Bonamia ostreae.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00018/12880/9829.pdf
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Molecular detection and quantification of the protozoan Bonamia ostreae in the flat oyster, Ostrea edulis ArchiMer
Robert, Maeva; Garcia, Celine; Chollet, Bruno; Lopez-flores, Inmaculada; Ferrand, Sylvie; Francois, Cyrille; Joly, Jean-pierre; Arzul, Isabelle.
Bonamia ostreae is an intracellular protozoan which is recognized as a cause of mortality in European populations of flat oysters (Ostrea edulis). Based on the recent characterization of actin genes of B. ostreae, specific primers were designed for real-time PCR using SYBR® Green chemistry. Specificity was demonstrated by the unique melting temperature peak observed in positive samples and by the lack of amplification in samples of oysters infected by closely related parasites, including Bonamia exitiosa. A calibration curve using a cloned template was defined to estimate copy number. The assay had a 6 log- dynamic range, mean inter- and intra-assay variation coefficients of <1% and a minimum detection limit of 50 gene copies per reaction. Using...
Tipo: Text Palavras-chave: Heart imprints; Ostrea edulis; Quantification; Detection; Real time PCR; Bonamia ostreae.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6934.pdf
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Morphology and expression of genes related to skeletal muscle growth in juveniles of pirarucu (Arapaima gigas, Arapaimatidae, Teleostei) - doi: 10.4025/actascianimsci.v35i3.18219 Animal Sciences
Carani, Fernanda Regina; Universidade Estadual Paulista (UNESP); Duran, Bruno Oliveira da Silva; Universidade Estadual Paulista (UNESP); Paula, Tassiana Gutierrez de; Universidade Estadual Paulista (UNESP); Piedade, Warlen Pereira; Universidade Estadual Paulista (UNESP); Dal-Pai, Maeli; Universidade Estadual Paulista (UNESP).
Skeletal muscle growth in the pirarucu (Arapaima gigas) is highly interesting to fish farmers because it provides information about how the mechanism in muscle mass increase, characteristic of the species, is regulated. Pirarucu has specific muscle growth that highlights the species’s significance and commercial value. Current research evaluates the morphology and the growth-related gene expression in the red and white skeletal muscles of the pirarucu. Muscle samples were collected from the lateral anterior region and frozen in liquid nitrogen. Histological sections were performed and stained by HE for morphological analysis. Red and white muscle samples were used to determine MyoD, myogenin, and myostatin genes expression by Real-time Polymerase Chain...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 5.06.03.00-0 muscle fibre morphology; Myogenic regulatory factors; Myostatin; Skeletal muscle growth; Arapaima gigas; Real time PCR.
Ano: 2013 URL: http://periodicos.uem.br/ojs/index.php/ActaSciAnimSci/article/view/18219
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Mycobacterium avium subsp. Paratuberculosis and the expression of selected virulence and pathogenesis genes in response to 6°c, 65°c and ph 2.0 BJM
Pribylova,Radka; Kralik,Petr; Donnelly,Neysan; Matiasovic,Jan; Pavlik,Ivo.
The aim of this work was to study the expression of selected Mycobacterium avium subsp. paratuberculosis (MAP) genes connected with MAP virulence, adhesion and stress response. The temperature of 6°C and 65°C were chosen with regard to the food industry, storage conditions (refrigerator) and low-temperature pasteurization. A pH of 2.0, using lactic acid, was selected to mimic the natural environment of the stomach. Expression of selected genes was studied using real time reverse transcription PCR on three different MAP isolates. MAP isolates were chosen according to the number of their preceding cultivations. While isolates 8672 and 8819 were previously cultivated only once, MAP isolate 12146 went through four passages. Different expression profiles were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: MAP; Johne's disease; Stress; Treatment; Real time PCR.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000200049
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Ostreid herpes virus 1 infection in families of the Pacific oyster, Crassostrea gigas, during a summer mortality outbreak: Differences in viral DNA detection and quantification using real-time PCR ArchiMer
Sauvage, Christopher; Pepin, Jean-francois; Lapegue, Sylvie; Boudry, Pierre; Renault, Tristan.
Ostreid herpes virus 1 (OsHV-1) infections, notably reported in Europe and the USA, are closely associated with significant mortalities of the Pacific oyster, Crassostrea gigas, especially during its early stages of life. In summer 2006, we monitored mortality by strict daily verification of three full-sib families of oysters reared under common conditions. We quantified OsHV-1 using real-time PCR in dead and living individuals during and after a mortality event. Mortality events were severe and brief, but significantly different between tested families (cumulative mortality ranging from 1.2 to 49%). Real-time PCR assays revealed different viral DNA loads in dead individuals from different families (P < 0.001). Moreover, the mean level of infection...
Tipo: Text Palavras-chave: Viral DNA quantification; Real time PCR; OsHV 1; Mortality; Crassostrea gigas.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6265.pdf
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Quantification of Vibrio penaeicida, the etiological agent of Syndrome 93 in New Caledonian shrimp, by real-time PCR using SYBR Green I chemistry ArchiMer
Goarant, Cyrille; Merien, F.
Shrimp farming is a small but growing industry in New Caledonia. Since 1993, "Syndrome 93" has been affecting New Caledonian shrimp farming industry every cold season, causing severe epizootic mortalities in grow-out ponds and significant losses. Highly pathogenic strains of Vibrio penaeicida are considered the etiological agent of the disease in Litopenaeus stylirostris. On one hand, studies demonstrated that healthy shrimp may carry V penaeicida for weeks with a high overall prevalence, regardless of any seasonal pattern or temperature conditions. On the other hand, larvae are free of V penaeicida and are also resistant to experimental infection. V penaeicida is frequently detected in incoming water pumped from the bays, which was shown, by a molecular...
Tipo: Text Palavras-chave: Vibriosis; Vibrio; Real time PCR; Quantification; Mariculture; Extraction techniques.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-1903.pdf
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