976 resultados para sequence characterized amplified region
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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The objective of this study was to evaluate the reproductive performance of Santa Ines rams subjected to successive semen collections in an Amazonian climate. Four rams were subjected to successive ejaculations during a maximum period of three hours. This procedure was repeated three times at 15-day intervals. Sexual and behavioural (libido) and andrological (testicular and seminal) assessments were performed. A total of 81 ejaculates were collected. Libido and semen vigour, volume, appearance and concentration decreased as the ejaculation frequency increased (P<0.05) and sperm motility showed a decreasing trend (P=0.06). The seminal pH increased over the sequence of collections (P<0.05). The only significant differences observed between individual rams were the variable scrotal circumference and the percentages of live sperm and sperm abnormalities (P<0.05). All the parameters of the first ejaculation were within the normal range for this species, which suggests that the local climatic conditions (high temperature and humidity) did not affect the behavioural, testicular or seminal parameters of experimental rams. Our findings indicate that the reproductive performance of Santa Ines rams could be affected by the intensification of ejaculation frequency; however, individual male variation needs to be taken into consideration.
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A semi-nested reverse transcription-polymerase chain reaction (Semi-N-RT-PCR) was developed and used to detect the S glycoprotein gene of infectious bronchitis virus (IBV) strains and to discriminate H120 vaccine strain from other strains. Viral RNA was extracted from the allantoic fluid of chicken embryos and from tissues of chickens experimentally infected with different strains of IBV. Amplification and identification of the viral RNA was performed using two sets of primers complementary to a region of the S glycoprotein gene in the Semi-N-RT-PCR assay. The pair of primers used in the first PCR consisted of universal oligonucleotides flanking a more variable region of S1-S2 gene. The second primer pair was used in the Semi-N-RT-PCR and was comprised of one of the primers from the first universal pair together with either another universal internal oligolucleotide or a oligonucleotide sequence specific for the H120 strain of IBV. The universal primers detected all reference IBV strains and field isolates tested herein. The Semi-N-RT-PCR had high sensitivity and specificity, and was able to differentiate the H120 vaccine strain from other reference IBV strains; including M41 strain. All tissue samples collected from chickens experimentally infected with H120 or M41 strains were positive in the semi-nested RT-PCR using universal primers, while only the H120-infected tissue samples were amplified by the set of primers containing the H120-oligonucleotide. In conclusion, the ability of Semi-N-RT-PCR to detect distinct IBV strains and preliminarily discriminate the vaccine strain (H120) closes a diagnostic gap and offers the opportunity to use comprehensive PCR procedures for the IBV diagnosis.
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LMV-Common and LMV-Most are two seed-borne types of Lettuce mosaic virus (LMV), genus Potyvirus. LMV-Most, but not LMV-Common, overcomes the resistance afforded to lettuce by two recessive genes, mo1(1) and mo1(2). An RT-PCR-based assay thought to be specific for LMV-Most also amplified LMV-Tn2, previously typified as LMV-Common. The sequence of selected regions along the genome indicated that LMV-Tn2 is a natural recombinant between LMV-Most and LMV-Common isolates, with a putative recombination site located within the P3 coding region. This is the first evidence of a naturally occurring LMV recombinant isolate.
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Paracoccidioides brasiliensis isolates are not homogeneous in their patterns of pathogenicity in animals and adhesion to epithelial cells. During this investigation, genotypic differences were observed between two samples of P. brasiliensis strain 18 yeast phase (Pbl 8) previously cultured many times, one taken before (Pb18a) and the other after (Pb18b) animal inoculation. Random amplified polymorphic DNA analysis using the primer OPJ4 distinguished Pb18b from Pbl Ba by one 308 bp DNA fragment, which after cloning and sequencing was shown to encode a polypeptide sequence homologous to the protein beta-adaptin. It is suggested, by comparison to other micro-organisms, that this protein might play an important role in the virulence of P. brasiliensis. This result demonstrates the influence of in vitro subculturing on the genotype of this organism.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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In a previous paper, the current state of knowledge of the region containing the Phocaea dynamical family was revised. Here, the dynamical evolution and possible origin of the Phocaea dynamical family and asteroid groups in the region are investigated. First, I study the case of asteroids at high eccentricity (e > 0.31). I find that these objects are unstable because of encounters with Mars on time-scales of up to 270 Myr. The minimum time needed by members of the Phocaea classical family to reach the orbital locations of these objects, 370 Myr, can be used to set a lower limit on the age of the Phocaea family.Next, attention is focused on the chaotic layer previously identified near the nu(6) secular resonance border. Using analytical and numerical tools, I find that the presence of the nu(6) secular resonance forces asteroids with vertical bar g-g(6)vertical bar < 2.55 arcsec yr(-1) to reach eccentricities high enough to allow them to experience deep, close encounters with Mars. Results of the analytical model of Yoshikawa and of my numerical simulations fully explain the low-inclination chaotic region found by Carruba.Finally, I investigate the long-term stability of the minor families and clumps identified in the previous paper, with particular emphasis on a clump only identifiable in the domain of proper frequencies (n, g, g - s) around (6246) Komurotoru. I find that while the clumps identified in the space of proper elements quickly disperse when the Yarkovsky effect is considered, the family around (19536) is still observable for time-scales of more than 50 Myr. The (6246) clump, characterized by its interaction with the nu(5) + nu(16) and 2 nu(6) - nu(16) secular resonances, is robust on time-scales of 50 Myr. I confirm that this group may be the first clump ever detected in the frequency domain that can be associated with a real collisional event.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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This thesis aims to advance in the geological knowledge of the region comprising the Piancó-Alto Brígida (TPAB) and Alto pajeú (TAP) terranes, in the Transversal Zone Domain (Borborema Province, NE Brazil), with the main objective of understanding the geodynamic evolution and the structural framework of these units. To reach this objective, and besides field work and interpretation of traditional aerial photographs, other tools were employed like of remote sensing products (Landsat 7 ETM+, aeroradiometrics, aeromagnetics and topographical images), lithogeochemical (whole rock) analyses and geochronological dating (U-Pb in zircon), besides integration with literature data. In the area, several precambrian geological units outcrop, represented in the TAP by the paleoproterozoic Serra Talhada and Afogados da Ingazeira complexes, Riacho Gravatá Complex (metavolcano-sedimentary sequence of Stenian-Tonian age) and Cariris Velhos orthogneisses (of Tonian age). The TPAB comprises the Santana do Garrote (lower unit) and Serra do Olho d'Água (upper unit) formations of the Cachoeirinha Group (Neoproterozoic III), besides the Piancó orthogneisses and Bom Jesus paragneisses; the latter correspond to an older (basement ?) block and a possible high grade equivalent of the Cachoeirinha Group (or Seridó Group ?), respectively. Several Brasiliano-age plutons occur in both terranes.The aeromagnetic data show the continuity, at depth, of the main shear zones mapped in the region. The Patos, Pernambuco, Boqueirão dos Cochos, Serra do Caboclo, Afogados da Ingazeira/Jabitacá and Congo-Cruzeiro do Nordeste shear zones reach depths greater than to 6-16 km. The aeromagnetic signature of other shear zones, like the Juru one, suggests that these structures correspond to shallower crustal features. The satellite images (Landsat 7 ETM+) and aerogamaspectrometric images discriminate different geological units, contributing to the mapping of the structural framework of the region. The Serra do Caboclo Shear Zone was characterized as the boundary/suture between the TPAB and TAP. This structure is an outstanding, pervasive feature that separates contrasting geological units, such as the Neoproterozoic III Cachoeirinha Group in the TPAB and the Riacho Gravatá Complex and the Cariris Velhos metaplutonics, of Stenian-Tonian age, in the TAP. Occupying different blocks, these units are not found in authoctonous relations, like unconformities and intrusive contacts. Concerning the Cariris Velhos (ca. 1,0 Ga old) event is recorded by radiometric ages of the Riacho Gravatá Complex metavolcanics and intrusive augen and orthogneisses, all of them displaying geochemical affinities of arc or collisional settings. A structural signature of this event was not recorded in the region, possibly due to its low grade/low strain style, obliterated by the overprinting of younger, higher grade/high strain Brasiliano-age fabrics.The first tectonic event (D1) observed in the Cariris Velhos lithotypes presents contractional kinematics with transport to the NW. Neoproterozoic III geochronologic dates, obtained in late-D1 granitoids, imply a Brasiliano age (ca. 610-600 Ma) for this deformation event. The second tectonic event (D2) characterized in the region corresponds to the Brasiliano transcurrent kinematics of the outstanding shear zones and associated granitoid plutons. The geochronological (U-Pb in zircon) data obtained during this thesis also confirms the occurrence of the Cariris Velhos magmatic suite in the TAP, as well as the Neoproterozoic III age to the Cachoeirinha Group in the TPAB. The TAP (Riacho Gravatá Complex, augen and orthogneisses) is interpreted as a continental arc possibly accreted to a microcontinent during the Cariris Velhos (Stenian-Tonian) event. Later on, this terrane collided with the TPAB at the beginning of the Brasiliano orogeny (D1 contractional deformation), and both domins were reworked by the transcurrent shear deformation of the D2 event
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The genome of all organisms is subject to injuries that can be caused by endogenous and environmental factors. If these lesions are not corrected, it can be fixed generating a mutation which can be lethal to the organisms. In order to prevent this, there are different DNA repair mechanisms. These mechanisms are well known in bacteria, yeast, human, but not in plants. Two plant models Oriza sativa and Arabidopsis thaliana had the genome sequenced and due to this some DNA repair genes have been characterized. The aim of this work is to characterized two sugarcane cDNAs that had homology to AP endonuclease: scARP1 and scARP3. In silico has been done with these two sequences and other from plants. It has been observed domain conservation on these sequences, but the cystein at 65 position that is a characteristic from the redox domain in APE1 protein was not so conservated in plants. Phylogenetic relationship showed two branches, one branch with dicots and monocots sequence and the other branch with only monocots sequences. Another approach in order to characterized these two cDNAs was to construct overexpression cassettes (sense and antisense orientation) using the 35S promoter. After that, these cassettes were transferred to the binary vector pPZP211. Furthermore, previously in the laboratory was obtained a plant from nicotiana tabacum containing the overexpression cassette in anti-sense orientation. It has been observed that this plant had a slow development and problems in setting seeds. After some manual crossing, some seeds were obtained (T2) and it was analyzed the T2 segregation. The third approach used in this work was to clone the promoter region from these two cDNAs by PCR walking. The sequences obtained were analyzed using the program PLANTCARE. It was observed in these sequences some motives that may be related to oxidative stress response
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)