997 resultados para degenerate primers
Resumo:
Es aquesta comunicació es presenten els primers resultats d’una recerca realitzada entre l’ICE de la UB i el GROP, i que s’ha portat a terme amb escoles que estan desenvolupant el Projecte Escolta’m. La finalitat és valorar la influencia del Projecte, que té com a base la tutoria personalitzada, en el desenvolupament de competència emocional dels alumnes que hi participen. Es van escollir 6 escoles que duen a terme el projecte, tot i que no totes porten el mateix temps aplicant-lo. Les escoles que duen a terme l’Escolta’m són escoles amb diferents poblacions, realitats i projectes. Totes tenen en comú un interès especial per la tutoria individualitzada. Les dades obtingudes aporten evidencies significatives de que el projecte està ajudant al desenvolupament de la majoria de competències emocionals.
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The objective of this work was to characterize mandarin (Citrus spp.) germplasm from Southern Brazil by morphological and molecular analyses. Thirty seven cultivars from 34 distinct mandarin varieties were evaluated by morphological and agronomic traits of leaves, flowers and fruits, and by microsatellite markers. The morphological and agronomic characteristics suggested that almost all varieties can be produced for commercial use, and some, as the Satsuma variety, are recommended for breeding programs. Pooled DNA samples from 1-5 plants belonging to each cultivar were tested. Eight of the nine primers detected polymorphisms. Specific markers were found for some accessions. The dendrogram constructed with the morphological results divided the 37 cultivars into four groups, while that obtained with the microsatellites clustered 35 of the 37 cultivars into three groups only. Generally, intervarietal differences are not high, and this lack of agreement in the two multifactorial analyses indicates that diverse evolutionary factors are acting at these two levels of investigation.
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Even though much improvement has been made in plant transformation methods, the screening of transgenic plants is often a laborious work. Most approaches for detecting the transgene in transformed plants are still timeconsuming, and can be quite expensive. The objective of this study was to search for a simpler method to screen for transgenic plants. The infiltration of kanamycin (100 mg/mL) into tobacco leaves resulted in conspicuous chlorotic spots on the non-transgenic plant leaves, while no spots were seen on the leaves of transformed plants. This reaction occurred regardless of age of the tested plants, and the method has proven to be simple, fast, non-destructive, relatively cheap, and reliable. These results were comparable to those obtained by the polymerase chain reaction (PCR) amplification of the transgene using specific primers.
Resumo:
O objetivo deste trabalho foi a caracterização genética da raça bovina Crioulo Lageano por marcadores RAPD em comparação com as raças Holandesa e Nelore. Foram selecionados 43 primers, que geraram 77 bandas polimórficas. Os animais foram distribuídos em cinco subgrupos de Crioulo Lageano (I a V), e um subgrupo em cada uma das raças Holandesa (VI) e Nelore (VII). A maior parte da variância genética total (65,05%) foi causada pela diferença de indivíduos dentro dos grupos, e o restante pelas diferenças entre grupos. A análise conjunta dos grupos I a V apresentou variabilidade genética entre grupos de 25,28% e dentro dos grupos de 74,72%. A diversidade gênica vem se mantendo ao longo das gerações no núcleo de conservação do Crioulo Lageano. A raça Holandesa apresentou a menor diversidade gênica (0,1204), e a Crioulo Lageano a maior (0,3154). A maior distância genética (0,3747) foi entre as raças Nelore e Holandesa. Os grupos de Crioulo Lageano apresentaram diferenças entre si e apenas alguns indivíduos de cada grupo posicionaram-se junto a outros grupos. A técnica RAPD é capaz de estimar a distância genética entre raças ou populações e de auxiliar na escolha de indivíduos, visando aos trabalhos de conservação de recursos genéticos.
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The objective of this study was to characterize the Peruvian isolate of Metarhizium anisopliae var. acridum, CG 863, obtained from the grasshopper Schistocerca interrita, a crop pest in Peru. The characterization was done by comparing this isolate with two other ones of M. anisopliae var. acridum, from Brazil and Australia, and with an isolate of M. anisopliae var. anisopliae. The three M. anisopliae var. acridum isolates had similar growth profiles in agar plates at 25°C and 37°C, and similar RAPD patterns according to the analysis of three primers. However, regarding these parameters and conidial size, these isolates were very distinct when compared to M. anisopliae var. anisopliae isolate. Bioassays indicated that the Peruvian isolate is as pathogenic as the Brazilian isolate against nymphs of Rhammatocerus schistocercoides.
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In vitro regeneration of Arachis retusa was examined for the purpose of germplasm renewal and conservation. Random amplified polymorphic DNA (RAPD) fingerprinting was used to evaluate the genetic stability of plants derived from embryo axes and apical segments. Ten arbitrary decamer primers were screened and five of them were selected. Ninety genomic regions were evaluated, with an average of 18 loci per clone. All amplified segments were monomorphic. The results indicate that recovered plants are genetically stable at the assessed genomic regions and that both regeneration processes are suitable for in vitro germplasm preservation of Arachis species.
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Conservation and improvement strategies should be based on the association between genetic and phenotypic characteristics. The objective of this work was to characterize five native Brazilian cattle breeds (Caracu, Crioulo Lageano, Curraleiro, National Polled and Pantaneiro) and two commercial breeds (Holstein and Nellore) using RAPD technique to estimate genetic distances and variability between and within breeds. Genetic relationships were investigated using 22 primers which generated 122 polymorphic bands. Analysis of molecular variance indicated that most of the genetic variation lay among individuals within populations. The genetic variabilities between pairs of breeds were statistically significant. The smallest genetic divergence was between Crioulo Lageano and Curraleiro.The National Polled, although historically considered to be of Bos taurus aquitanicus origin,similar to theCaracu, was grouped together with the other breeds of Bos taurus ibericus origin. Generally, the individual breeds formed distinct clusters except the National Polled. The RAPD technique was capable to distinguish genetically between the breeds studied; the Caracu, Crioulo Lageano, Curraleiro and Pantaneiro may be considered distinct genetic entities thereby proving the uniqueness of the populations; the National Polled has not been able to re-establish itself after its decline in the 1950s, thereby losing its genetic identity.
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The objectives of this work were to evaluate the frequency of polyembryony, and to identify zygotic and nucellar seedlings of Citrus volkameriana using RAPD. Twenty-five polyembryonic and eight monoembryonic seeds were cultivated in vitrofor six months. DNA from seedlings was extracted and used in combination with five RAPD primers to identify zygotic or nucellar origin of the seedlings. Environmental conditions of the year affected significantly (P<0.05) the morphological characteristics of fruitsand the number ofembryos per seed. Polyembryonic seeds ranged from 30.9%, 44.8% to 54.4% over three years. Morphological characteristic was not correlated with polyembryony. In vitro culture enable all embryos of each seed to grow, favoring the percentage of seedlings identified as zygotic. In polyembryonic and monoembryonic seeds, 25.9% and 87.5% of the seedlings, respectively, were sexually originated. In polyembryonic seeds, not all zygotic seedlings were produced by small embryos located at the micropyle.
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The objective of this work was to determine the genetic differences among eight Brazilian populations of the tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), from the states of Espírito Santo (Santa Tereza), Goiás (Goianápolis), Minas Gerais (Uberlândia and Viçosa), Pernambuco (Camocim de São Félix), Rio de Janeiro (São João da Barra) and São Paulo (Paulínia and Sumaré), using the amplified fragment length polymorphism (AFLP) technique. Fifteen combinations of EcoRI and MseI primers were used to assess divergence among populations. The data were analyzed using unweighted pair-group method, based on arithmetic averages (UPGMA) bootstrap analysis and principal coordinate analysis. Using a multilocus approach, these populations were divided in two groups, based on genetic fingerprints. Populations from Goianápolis, Santa Tereza, and Viçosa formed one group. Populations from Camocim de São Félix, Paulínia, São João da Barra, Sumaré, and Uberlândia fitted in the second group. These results were congruent with differences in susceptibility of this insect to insecticides, previously identified by other authors.
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Garlic viruses often occur in mixed infections under field conditions. In this study, garlic samples collected in three geographical areas of Brazil were tested by Dot-ELISA for the detection of allexiviruses using monoclonal specific antibodies to detect Garlic virus A (GarV-A), Garlic virus B (GarV-B), Garlic virus C (GarV-C) and a polyclonal antiserum able to detect the three virus species mentioned plus Garlic virus D (GarV-D). The detected viruses were biologically isolated by successive passages through Chenopodium quinoa. Reverse Transcriptase Polimerase Chain Reaction (RT-PCR) was performed using primers designed from specific regions of the coat protein genes of Japanese allexiviruses available in the Genetic Bank of National Center of Biotechnology Information (NCBI). By these procedures, individual garlic virus genomes were isolated and sequenced. The nucleotide and amino acid sequence analysis and the one with serological data revealed the presence of three distinct allexiviruses GarV-C, GarV-D and a recently described allexivirus, named Garlic mite-borne filamentous virus (GarMbFV), in Brazil.
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The objective of this work was to characterize 27 potato genotypes, using molecular markers. Polyacrylamide gel electrophoresis, RAPD techniques and isozymes of esterase, phosphoglucomutase and soluble proteins were analyzed in tubers, and isocitrate dehydrogenase, aspartate transaminase, phosphoglucomutase and peroxidase, in leaves. Eighteen primers were tested and four were chosen, kits OPX (01, 04 and 09) and OPY (07), to analyze RAPD markers in leaf extracts. Similarity and cluster analysis were conducted using Jaccard coefficient and the unweighted pair-group method using arithmetic average. Despite the differences detected in the analysis of proteins and isozymes in the tubers, as well as of isozymes in the leaves, the characterization of all genotypes through gel electrophoresis was not possible, while RAPD markers were efficient to characterize all the 27 genotypes.
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La manera en que el triatleta és capaç d’adaptar-se a la cursa a peu després de la segona transició, és decisiva en el resultat final en una prova de triatló. En aquest estudi s’avalua l’efecte del ciclisme sobre la longitud i freqüència de gambada durant la simulació de una transició ciclisme-carrera de traiatló en esportistes sub 23 i juniros élite. Cinc subjectes van realitzar dues proves: 1) Carrera de control (C), que consisteix en 3000 metres de carrera a màxima velocitat en una pista de 400 m (7’5 voltes). 2) Transició (T), consisteix en 30’ de ciclisme, 10’ d’escalfament i 20’ a zona de llindar anaeròbic, seguits de 3000 m de carrera a màxima velocitat possible. Es van mostrar diferencies importants amb la longitud de gambada: 339,61 cm a la prova C i 302,74 cm en la prova T, sobre tot als primers 600 metres, i en cap moment es va arribar a la longitud de la cursa C. Per tant concloem, que en la T2 hi ha un efectes negatius sobre la longitud de gambada durant els 3000 m, però sobretot, als primer 600 metres.
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The objective of this study was to verify the genetic diversity between and within seven populations of Moxotó goat (n = 264) from the States of Pernambuco, Paraíba and Rio Grande do Norte, using RAPD (Random Amplified Polymorphic DNA). Moxotó, as well as other naturalized breeds, suffers genetic losses due to the indiscriminate miscegenation with breeds raised in the Northeast Region of Brazil. The genetic characterization of these genetic resources is essential to conservation and breeding programs. DNA was extracted from lymphocytes using a non-organic protocol. The 16 primers used were selected from 120 decamer oligonucleotide primers and generated 56 polymorphic bands. The analysis of molecular variance (AMOVA) showed that the greater part of total genetic variability (71.55%) was due to differences between individuals within populations, while 21.21% was among populations. The analysis of variance among the pairs of populations demonstrated that the populations located in Floresta, PE x Angicos, RN presented a smaller value of intrapopulational differentiation (8.9%), indicating low genetic variability among them. Nei's genetic distances varied between 0.0546 and 0.1868 in the populations. The dendrogram generated showed that the Canindé breed, used as outgroup, clustered with the populations of Moxotó, indicating a possible common origin of the naturalized goat breeds.
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Pollination in flowering plants requires that anthers release pollen when the gynoecium is competent to support fertilization. We show that in Arabidopsis thaliana, two paralogous auxin response transcription factors, ARF6 and ARF8, regulate both stamen and gynoecium maturation. arf6 arf8 double-null mutant flowers arrested as infertile closed buds with short petals, short stamen filaments, undehisced anthers that did not release pollen and immature gynoecia. Numerous developmentally regulated genes failed to be induced. ARF6 and ARF8 thus coordinate the transition from immature to mature fertile flowers. Jasmonic acid (JA) measurements and JA feeding experiments showed that decreased jasmonate production caused the block in pollen release, but not the gynoecium arrest. The double mutant had altered auxin responsive gene expression. However, whole flower auxin levels did not change during flower maturation, suggesting that auxin might regulate flower maturation only under specific environmental conditions, or in localized organs or tissues of flowers. arf6 and arf8 single mutants and sesquimutants (homozygous for one mutation and heterozygous for the other) had delayed stamen development and decreased fecundity, indicating that ARF6 and ARF8 gene dosage affects timing of flower maturation quantitatively.
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The peroxisome proliferator-activated receptor gamma (PPARgamma) mediates the activity of the insulin-sensitizing thiazolidinediones and plays an important role in adipocyte differentiation and fat accretion. The analysis of PPARgamma functions in mature adipocytes is precluded by lethality of PPARgamma(-/-) fetuses and tetraploid-rescued pups. Therefore we have selectively ablated PPARgamma in adipocytes of adult mice by using the tamoxifen-dependent Cre-ER(T2) recombination system. We show that mature PPARgamma-null white and brown adipocytes die within a few days and are replaced by newly formed PPARgamma-positive adipocytes, demonstrating that PPARgamma is essential for the in vivo survival of mature adipocytes, in addition to its well established requirement for their differentiation. Our data suggest that potent PPARgamma antagonists could be used to acutely reduce obesity.