Made available in DSpace on 2015-04-22T22:12:36Z (GMT). No. of bitstreams: 1 carolinie.pdf: 1105289 bytes, checksum: fb4a9af7cf4012e7784dfbcf92246455 (MD5) Previous issue date: 2010-03-09 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / In the Amazon agricultural soils are found in soil called Anthropogenic Dark Earths (ADE), a soil rich in organic matter and minerals. The biodiversity is a source of very high value of non-cultivable microorganisms that can be isolated through construction of metagenomic libraries, using vectors of large fragments, with the goal of finding new biocatalytic enzymes. Enzymes widely used in the industrial market are amylase and protease, are applied in industry: chemical, pharmaceutical, textile, detergent and food industries. Amylases degrade starch into smaller units of saccharides, as proteases catalyze the hydrolysis of peptide bonds. The aim of this study was to isolate and partially characterize enzymes amylase and protease of clones isolated from a selection of functional metagenomic library of TPI in the Amazon region. In this study were isolated 1.344 clones, only 4 producing halo of starch hydrolysis and 3 proteolytic clones belonging to metagenomic library constructed with total DNA extracted from microorganisms present in soil samples of the TPI, were partially characterized the enzymes produced by 4 clones amilolytic and 1 proteolytic clone. After isolation, the clones were inoculated in culture medium Luria Bertani (LB) at 37ºC under agitation of 150 rpm and subjected to assays at pHs of 3 to 10 and temperatures ranging from 25 to 100ºC for up to 90 of incubation. Amylases had its highest production in 24 h (4.3 U/mL) and its activity was optimal at neutral pH to slightly alkaline, the clone P1C4 showed the highest production at 70°C (6.93 U/mL), since the clones P5C4 and P6C12 reported higher thermostability at 80°C, these results indicate that the gene for alpha-amylase to be cloned encoding a thermostable enzyme, such as the enzyme produced by Bacillus liqueniformis. Since the clone P3A3 produced 26.3 U/mL protease in 10 hours of production, its activity was optimal at neutral pH, and its optimum temperature at 30ºC (56.0 U/mL) and its thermostability was demonstrated in 50°C (22,0 U/mL). These features contribute to the implementation of these enzymes in industrial sectors as in food production, chemical industry and production of detergents. The enzymes isolated in this study demonstrated new features and performance when compared to enzymes described today, which proves the efficiency of the construction of metagenomic libraries for isolation of new bioproducts. / Na floresta Amazônica são encontrados solos agricultáveis denominado de solo de Terra Preta de Índio (TPI), um solo rico em matéria orgânica e em minerais. A biodiversidade da Amazônia constitui uma fonte de valor altíssimo de micro-organismos não-cultiváveis que podem ser isolados através das construções das bibliotecas metagenômicas, utilizando vetores de grandes fragmentos, com o objetivo de encontrar novas enzimas biocatalíticas. As enzimas de maior aplicação no mercado industrial são as amilases e proteases, aplicadas nas indústrias: química, farmacêutica, têxtil, de detergentes e alimentícia. As amilases são capazes de degradar o amido em unidades de sacarídeos menores, já as proteases catalisam a hidrólise de ligações peptídicas. O objetivo deste trabalho foi isolar e caracterizar parcialmente enzimas amilolíticas e proteolíticas de clones isolados a partir de uma seleção funcional da biblioteca metagenômica de TPI da região Amazônica construída com DNA total extraído dos micro-organismos presentes nas amostras de solos de TPI, inseridos no vetor fosmidial (pCC1Fos ) e clonado na célula hospedeira Escherichia coli (EPI300). De 1.344 clones pertencente a biblioteca metagenômica, apenas 4 clones foram produtores de halo de hidrólise de amido e 3 clones foram proteolíticos, além disso, todas as enzimas produzidas pelos 4 clones amilolíticos e 1 clone proteolítico foram caracterizadas parcialmente. Após o isolamento, os clones foram inoculados em meio de cultura Luria Bertani (LB) a 37oC, sob agitação de 150 rpm e submetidos aos ensaios enzimáticos nos pHs de 3 a 10 e temperatura variando de 25 a 100ºC por até 90 minutos de incubação. As amilases tiveram sua maior produção em 24 horas (4,3 U/mL) e sua atividade ótima foi em pH neutro a levemente alcalino, o clone P1C4 demonstrou a maior produção a 70ºC (6,93 U/mL), já os clones P5C4 e P6C12 demonstraram a maior termoestabilidade a 80°C, tais resultados indicam que o gene de alfa-amilase clonado deve ser codificador de uma enzima termoestável, como por exemplo, a enzima produzida por Bacillus liqueniformis. Já o clone P3A3 produziu 26,3 U/mL de proteases em 10 horas de produção, sua atividade ótima foi em pH neutro, sendo sua temperatura ótima em 30ºC (56,0 U/mL) e sua termoestabilidade foi demonstrada em 50ºC (22,0 U/mL). Estas características contribuem para a aplicação destas enzimas em setores industriais como na produção de alimentos, indústria química e produção de detergentes. As enzimas isoladas neste trabalho demonstraram características e atuações novas quando comparadas a enzimas descritas atualmente, a qual comprova a eficiência da construção de bibliotecas metagenômicas para o isolamento de novos bioprodutos.

The wide diversity of plant secondary products results from different modifications undergone during biosynthesis, including glucosylation. These modification reactions result in production of the compounds actually found in plants and to unique chemical and biochemical properties, including some bitter compounds in grapefruit. While the presence of a PSPG box motif allows for identification of a clone as a putative glucosyltransferase (PGT), diversity of GT primary structures makes it difficult to accurately assign specific function. Our approach is to identify and isolate putative GT clones, express them heterologously, and biochemically characterize the proteins. Eleven putative GT clones have been isolated from Citrus paradise and some have been biochemically characterized. The current hypothesis being tested is that PGT3 and PGT9 clones are plant secondary product GTs. Due to issues with inclusion bodies when using E. coli, proteins were expressed in Pichia pastoris using the pPICZA vector. Recombinant protein expression was confirmed by Western blot and proteins were enriched by IMAC. Over 30 flavonoid and simple phenolic substrates, representing many compounds found in grapefruit, were screened for activity with PGT3 and PGT9 proteins. No significant activity was found and the biochemical function of the proteins encoded by these clones will be further investigated.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2009. / Härtill 5 uppsatser.

Software forking---creating a variant product by copying and modifying an existing project---is often considered an ad hoc, low cost alternative to principled product line development. To maintain forked projects, developers need to manually port existing features or bug-fixes from one project to another. Such manual porting is not only tedious but also error-prone. When the contexts of the ported code vary, developers often have to adapt the ported code to fit its surroundings. Faulty adaptations or inconsistent updates of the ported code could potentially introduce subtle inconsistencies in the codebase. To build a deeper understanding to cross-system porting and porting related errors, this dissertation investigates: (1) How can we identify ported code from software version histories? (2) What is the overhead of cross-system porting required to maintain forked projects? (3) What is the extent and characteristics of porting errors that occur in practice? and (4) How can we detect and characterize potential porting errors? As a first step towards assessing the overhead of cross-system porting, we implement REPERTOIRE, a tool to analyze repeated work of cross-system porting across peer projects. REPERTOIRE can detect ported edits between program patches with high accuracy of 94% precision and 84% recall. Using REPERTOIRE, we study the temporal, spatial, and developer dimensions of cross-system porting using 18 years of parallel evolution history of the BSD product family. Our study finds that cross-system porting happens periodically and the porting rate does not necessarily decrease over time. The upkeep work of porting changes from peer projects is significant and currently, porting practice seems to heavily depend on developers doing their porting job on time. Analyzing version histories of Linux and FreeBSD, we derive five categories of porting errors, including incorrect control- and data-flow, code redundancy, and inconsistent identifier and token renamings. Leveraging this categorization, we design a static control- and data-dependence analysis technique, SPA, to detect and characterize porting inconsistencies. SPA detects porting inconsistencies with 65% to 73% precision and 90% recall, and identify inconsistency types with 58% to 63% precision and 92% recall on average. In a comparison with two existing error detection tools, SPA outperforms them with 14% to 17% better precision. / text

Planar cell polarity (PCP) is the organization of cellular characteristics within the plane of a tissue. PCP manifests both structurally, as in the directionality of insect bristles or mammalian skin hair, or dynamically, as in vertebrate neurulation, gastrulation, and oriented cell division in the kidney. Two well-conserved pathways are known to regulate PCP in invertebrates and in vertebrates: the Frizzled/PCP pathway and the Fat-Dachsous (Ft-Ds) pathway. The latter consists of the cadherins Ft and Ds, along with the Golgi kinase Four-jointed (Fj) and the transcriptional co-repressor Atrophin (Atro). Ft and Ds can bind each other, suggesting a mechanism for signal transduction. Fj phosphorylates Ft and Ds, modulating their binding affinities for each other. Atro is proposed to link Ft-Ds signaling with downstream events in the nucleus during eye development. The details of Ft-Ds binding, and the consequences of their interactions with other members of the pathway are poorly understood. In this work, I quantitatively analyzed Ft-Ds pathway mutant clones for their effects on ommatidial polarity in the Drosophila eye. My findings suggest that the Ft-Ds pathway regulates PCP independently of asymmetric cellular accumulation of Ft or Ds. I found that Atro has a position-specific role in regulating polarity in the eye, that Fj dampens clonal polarity signals, and that asymmetric accumulation of the atypical myosin Dachs is not essential for production and propagation of a long-range PCP signal. My observations suggest that Ft and Ds interact to modulate a secondary signal that regulates long-range polarity, that signaling by the Ds intracellular domain is dependent on Ft, and that ommatidial fate specification is genetically separable from long-range signaling.

Darbo tikslas Nustatyti paprastosios pušies (Pinus sylvestris L.) klonų išsilaikymą ir fenologijos ypatumus lokalios amoniako taršos sąlygomis. Darbo objektas Tyrimai 2011 m. vykdyti Jonavos miškų urėdijos, Pagelažių girininkijos teritorijoje, esančiose sėklinėje plantacijoje, kuri nuo AB „Achemos yra nutolusi 6 km. Sėklinė plantacija buvo įveista 1966-1971 m. Pasodinta 30 vietinės kilmės pliusinių medžių klonų, naudota kvadratinė schema. Metodika Buvo išmatuoti visų medžių skersmenys Šiaurės-Pietų ir Vakarų-Rytų kryptimis bei įvertinta defoliacija. Apskaičiuoti visų medžių klonų vidurkiai, F kriterijus, bei paveldėjimo koeficientas. Duomenų analizei panaudota programa STATISTICA. Darbo rezultatai Tarp nagrinėjamų parametrų tarp atskirų klonų buvo esminiai genetiniai skirtumai. Paveldėjimo koeficientas − nuo 0.13 iki 0.18. Defoliacijos paveldėjimo koeficientas buvo didesnis negu skersmens. Išskirti atskiri klonai, kurių parametrai turi esminį skirtumą. Mažesnės defoliacijos klonai buvo didesnio skersmens. Darytina prielaida, kad šie klonai gerai toleruoja aplinkos taršą. Šiai hipotezei patvirtinti reikalinga klonų metinio radialinio prieaugio genetinio salygotumo tyrimas, skirtingose oro amoniako taršos koncentracijose. / Purpose The aim of the study was to determine retenton and phenology of Scots pine (Pinus sylvestris L.) clones under local ammonia pollution condition. Object Investigation was accomplised in seed plantation of Pageleziai forestry of Jonava forestry district distance 6 km from nitrogen fertilizer factory. Seed plantation was created in 1966-1971 m. There was planted 30 local genesis plus trees of Scots pine clones and exposition of field with quadratic scheme. Methods During investigation was measured all trees diametrical values of the North-South and West-East direction and the same valued defoliation. All trees were calculated means, F-test, and heritage coefficient. Describing data analysis used STATISTICA. Results Investigation between all parameters and individual clones assessed significant genetics differences. Heritage coefficient range between 0.13 and 0.18. Heritage coefficient of the defoliation were larger than diametrical values. Seperation of the individual clones parameters showed significant diferences. Clones with less defoliation were larger diameters. Assumption of this study is that these clones can tolerate environmental pollution. The hypothesis confirmed are needed annual radial increment of the clones study of the genetic conventionality under different concentration of air ammonia pollution.

Planar cell polarity (PCP) is the organization of cellular characteristics within the plane of a tissue. PCP manifests both structurally, as in the directionality of insect bristles or mammalian skin hair, or dynamically, as in vertebrate neurulation, gastrulation, and oriented cell division in the kidney. Two well-conserved pathways are known to regulate PCP in invertebrates and in vertebrates: the Frizzled/PCP pathway and the Fat-Dachsous (Ft-Ds) pathway. The latter consists of the cadherins Ft and Ds, along with the Golgi kinase Four-jointed (Fj) and the transcriptional co-repressor Atrophin (Atro). Ft and Ds can bind each other, suggesting a mechanism for signal transduction. Fj phosphorylates Ft and Ds, modulating their binding affinities for each other. Atro is proposed to link Ft-Ds signaling with downstream events in the nucleus during eye development. The details of Ft-Ds binding, and the consequences of their interactions with other members of the pathway are poorly understood. In this work, I quantitatively analyzed Ft-Ds pathway mutant clones for their effects on ommatidial polarity in the Drosophila eye. My findings suggest that the Ft-Ds pathway regulates PCP independently of asymmetric cellular accumulation of Ft or Ds. I found that Atro has a position-specific role in regulating polarity in the eye, that Fj dampens clonal polarity signals, and that asymmetric accumulation of the atypical myosin Dachs is not essential for production and propagation of a long-range PCP signal. My observations suggest that Ft and Ds interact to modulate a secondary signal that regulates long-range polarity, that signaling by the Ds intracellular domain is dependent on Ft, and that ommatidial fate specification is genetically separable from long-range signaling.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2002. / Härtill 5 uppsatser.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-05-05T15:56:30Z No. of bitstreams: 1 texto completo.pdf: 399359 bytes, checksum: ae716e36f544e5e13da15f5717608860 (MD5) / Made available in DSpace on 2017-05-05T15:56:30Z (GMT). No. of bitstreams: 1 texto completo.pdf: 399359 bytes, checksum: ae716e36f544e5e13da15f5717608860 (MD5) Previous issue date: 2002-04-30 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / As variedades de cana-de-açúcar têm sido obtidas tanto por meio de acasalamentos biparentais como por policruzamentos. Entretanto, são poucos os relatos na literatura sobre as estimativas de parâmetros genéticos e os resultados da seleção confrontando os diferentes sistemas de acasalamento. As sementes foram derivadas de 15 famílias provenientes dos acasalamentos biparentais, policruzamento e autofecundação e foram semeadas em casa de vegetação. Após aproximadamente 20 a 30 dias as plântulas foram transplantadas individualmente para campo espaçadas no sulco a 0,50m. As mudas para instalação do experimento foram obtidas na soca, em fevereiro de 2000. Neste mesmo mês, o experimento foi implantado no Campo Experimental Fundão, próximo ao aeroporto, pertencente à Universidade Federal de Viçosa. Cada unidade experimental foi constituída de um sulco de 2m de comprimento, espaçados 1,40m entre si. Em cada unidade, foram plantadas 36 gemas. O delineamento empregado foi o de blocos aumentados, com as variedades RB855536 e SP80-1816 como tratamentos comuns em todos os blocos. Foram avaliados os caracteres: número de colmos, peso médio de colmos, Brix, comprimento médio de colmos, diâmetro médio de colmos, tonelada de colmos por hectare, tonelada de Brix por hectare e produção estimada de colmos em kg. A partir das análises intrablocos, foram obtidas as médias ajustadas dos tratamentos regulares e essas foram utilizadas para compor duas novas análises de variância. O teste F foi significativo para todos os caracteres, indicando variabilidade genética entre os sistemas de acasalamento. As médias do acasalamento biparental não diferiram estatisticamente das médias do policruzamento. As médias das famílias obtidas por autofecundação para os caracteres Brix e número de colmos não apresentaram diferença acentuada em relação aos outros sistemas de acasalamentos. Já para os outros caracteres há evidências de elevada depressão endogâmica. As estimativas das variâncias genéticas e herdabilidades para os sistemas de acasalamentos foram de magnitudes semelhantes. A variância genética dentro de família representou grande parte da variância genética total para todos os caracteres, exceto para TCH, para o qual a variância genética entre famílias representou 78% da variância genética total. A herdabilidade da família foi maior do que a herdabilidade de clone para quase todos os caracteres. / The sugar cane varieties have been obtained both by biparental matings and by mid-parents. However, there are few reports in literature on the estimates of genetic parameters and the results of the selection confronting the different mating systalks. The seeds were derived from 15 coming families of the biparental matings, midparent and selfing and they were sowed in greenhouse. After approximately 20 to 30 days, the plantules were transplanted individually for field placed in the furrow with a half-meter space. The seedlings for installation of the experiment were obtained in a ratton in February 2000. In this same month, the experiment was implanted in “Campo Experimental Fundão”, close to the airport, belonging to the Universidade Federal de Viçosa. Each experimental unit was constituted of a furrow of 2m of length, spaced 1,40m amongst themselves. In each unit, 36 yolks were planted. The experimental design was augmented blocks with the varieties RB855536 and SP80-1816 as common treatments in all of the blocks. The characters evaluated were: number of stalks, average stalk weight, Brix, average stalk length , average stalk diameter, stalk tons for hectare, Brix tons for hectare and estimated stalk production in kg. The adjusted averages of the regular treatments were obtained from the intrablocks and were used to compound two new variance analyses. The F test was significant for all of the characters, indicating genetic variability among the mating systalks. The averages of the biparental mating didn't differ statistically from the midparent averages. The families’ averages obtained by selfing for the characters Brix and number of stalks didn't differ much in relation to the other mating systalks. However, for the other characters, there are evidences of high endogamic depression. The estimates of the genetic variances and heritability for the mating systalks had similar magnitudes. The genetic variance in the family represented a great part of the total genetic variance for all of the characters, except for TCH, for which the genetic variance among families represented 78% of the total genetic variance. The family heritability was larger than the clone one for almost all of the characters. / Não foi localizado o currículo lattes do autor.

No description available.