Caracterização funcional de dois cDNAs homólogos à ciclofilina e à proteína reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para floração em tomateiro

Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

Caracterização funcional de dois cDNAs homólogos à ciclofilina e à proteína reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para floração em tomateiro

Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

Caracterização de genes de reparo de DNA em cana-de-açúcar

Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Caracterização de genes de reparo de DNA em cana-de-açúcar

Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Análise de cDNAs identificados em bibliotecas substrativas de cDNA para floração de variedades de cana-de-açúcar cultivadas no Rio Grande do Norte(RN): fator de transição NAC, Calmodulina e Fosfatidiltransferase

The flowering is a physiological process that it is vital for plants. This physiological process has been well studied in the plant model Arabidopsis, but in sugarcane this process is not well known. The transition of the shoot apical meristem from vegetative to flowering is a critical factor for plant development. At Brazil northeastern region, the transition to flowering in sugarcane has an important effect as it may reduce up to 60% its production. This is a consequence of the sugar translocation from stalks to the shoot apical meristem which is necessary during the flowering process. Therefore, the aim of this work was to explore and analyze cDNAs previously identified using subtractive cDNA libraries. The results showed that these cDNAs showed differential expression profile in varieties of sugarcane (early x late flowering). The in silico analysis suggested that these cDNAs had homology to calmodulin, NAC transcription factor and phosphatidylinositol, a SEC14, which were described in the literature as having a role in the process of floral development. To better understand the role of the cDNA homologous to calmodulin, tobacco plants were transformed with overexpression cassettes in sense and antissense orientation. Plants overexpressing the cassette in sense orientation did not flowered, while plants overexpressing the cassette in the antissense orientation produced flowers. The data obtained in this study suggested the possible role from CAM sequence, SEC14 and NAC in the induction/floral development pathway in sugarcane, this is the first study in order to analyze these genes in the sugarcane flowering process.

Floração precoce em cana-de-açúcar - um estudo utilizando ferramentas de análise in silico e proteômica

Sugarcane is one of the most important products of the world and Brazil is responsible for 25 % of the world production. One problem of this culture at northeast of Brazil is the early flowering. In our laboratory, it has been made before four subtractive libraries using early and late flowering genotypes in order to identify messages related to the flowering process. In this work, two cDNAs were chosen to make in silico analysis and overexpression constructs. Another approach to understand the flowering process in sugarcane was to use proteomic tools. First, the protocol for protein extraction using apical meristem was set up. After that, these proteins were separated on two bidimensional gels. It was possible to observe some difference for some regions of these gels as well as some proteins that can be found in all conditions. The next step, spots will be isolated and sequence on MS spectrometry in order to understand this physiological process in sugarcane

Amplificação, clonagem e sequenciamento de promotores de café (Coffea arabica cv Mundo Novo) específicos de fruto e ubíquos

The Coffee Genome Project made available to the scientific community relevant information that made practical the identification and cloning of important genes, as well as the identification of the major sequences involved on their regulation. The aim of the present study was to amplify, clone and sequence coffee promoters with specific expression patterns. For that, coffee ESTs which known expression profiles were employed. First, the promoter regions of coffee genes showing, respectively, fruitspecific and ubiquitous expression were amplified using the Genome Walking strategy. Amplified sequences were then inserted in the pGEM-Teasy vector (Promega) and sequenced. Once completed the sequencing, an expression cassette was constructed using the binary vector pCAMBIA-1381z (Cambia). These expression cassettes were cloned into Agrobacterium tumefaciens, and transgenic tobacco plants were generated aiming the functional characterization of these promoters

Narratives of the 'everyday' in Rio de Janeiro's favelas : community photographers' attempts at generating alternative news values [Narrativas do cotidiano nas favelas do Rio de Janeiro: tentativas de construção de valores-notícia alternativos pelos fotógrafos populares]

Abstract This paper presents the partial results of an ongoing research project which investigates ethnographically community (photo)journalism media initiatives, in Rio de Janeiro, Brazil. The Viva Rio and Observatorio de Favelas (NGOs) run projects that provide favela residents with skills to take, edit and print their own (photo) journalism contents that enables community-based framing and documentation of favela live, personalities and issues. Three months of fieldwork related to these projects in Rio's favelas generated remarkable theoretical issues that represent the community photographers' attempt to establish counter news values by shifting the focus from poverty, shortages, violence and criminality to images of the ordinary life which included the myriad events that occurs in the day of the favelas. Resumo Este artigo apresenta os resultados parciais de uma pesquisa em andamento que, a partir do método etnográfico, investiga os projetos de comunicação comunitária, jornalismo e fotojornalismo, no Rio de Janeiro, Brasil. As organizações não-governamentais (ONGs) Viva Rio e Observatório de Favela apoiam projetos que objetivam tornar moradores de favelas capazes de produzir, editar e publicar as suas próprias narrativas sobre personagens e questões do cotidiano de suas comunidades. O trabalho de campo sobre estes projetos, realizado durante três meses nas favelas do Rio de Janeiro, forneceu questões teóricas relevantes que representam o esforço dos fotógrafos populares para estabelecer contra valores-notícia transferindo o foco da pobreza, escassez, violência e criminalidade para imagens do cotidiano que inclui uma miríade de eventos que acontecem no dia-a-dia das favelas.

A construção da democracia : síntese histórica dos grandes momentos da Câmara dos Deputados, das Assembléias Nacionais Constituintes do Congresso Nacional

Síntese histórica dos grandes momentos da Câmara dos Deputados, das Assembléias Nacionais Constituintes e do Congresso Nacional.

A construção do artigo 5º da Constituição de 1988

Estuda a construção do artigo 5º da Constituição e mostra a transformação do seu texto durante o processo legislativo constituinte.

A participação feminina na construção de um parlamento democrático

Analisa a participação feminina na conquista da cidadania brasileira perante a Assembleia Constituinte de 1933 e a Assembleia Nacional Constituinte de 1987. Os dados foram coletados nos volumes do Diário do Congresso Nacional de 1933 e no portal do Congresso Nacional - Constituição 20 anos. Descreve o que foi consolidado como direitos para as mulheres. O estudo concluiu que há uma discrepância entre a norma constitucional vigente e as práticas sociais na realidade da mulher brasileira.

Agrotóxicos - construção da legislação

Consultoria Legislativa - Área XVI - Saúde Pública, Sanitarismo.

O papel do Poder Legislativo na construção do Sistema Único de Saúde: a Frente Parlamentar da Saúde e a regulamentação da Emenda à Constituição n. 29

Analisa o comportamento da Frente Parlamentar da Saúde durante o ano de 2007, com ênfase na apreciação do Projeto de Lei Complementar nº 1 de 2003. Apresenta referencial teórico que contextualiza o processo de construção do Sistema Único de Saúde no Brasil e o papel do Poder Legislativo na definição das diretrizes para as políticas públicas no setor saúde.

SINAPI x ORSE : análise comparativa entre o Sistema Nacional de Pesquisa de Custos e Índices da Construção Civil e o sistema adotado pelo Governo do Estado de Sergipe

Apresenta os resultados de um estudo comparativo entre os serviços e preços de construção civil disponibilizados por dois sistemas de custos: Sistema Nacional de Pesquisa de Custos e Índices da Construção Civil - SINAPI e Sistema de Orçamento de Obras de Sergipe - ORSE. Este estudo de caso realizado a partir de pesquisa documental e bibliográfica em que foram comparados dois orçamentos para um mesmo projeto de casa térrea padrão popular do Programa Minha Casa Minha Vida - MCMV: um com custos unitários extraídos do SINAPI Referencial e o outro com custos unitários originários do ORSE, ambos para o município de Aracaju, referentes a setembro de 2009. Os resultados indicaram a diversidade de métodos utilizados na formulação de composições unitárias de serviços para uso na construção civil e obras públicas.

Identificando um modelo para a construção de catálogo de serviços de tecnologia da informação em uma típica organização brasileira

A entrega de serviços de tecnologia da informação de baixa qualidade ainda é uma realidade nas empresas brasileiras. Clientes e usuários frustrados e baixo alinhamento dos serviços com as funções vitais do negócio estão associadas diretamente a essa percepção de qualidade. Estuda um dos primeiros passos a ser dado na busca de elevação da maturidade na entrega desses serviços: a construção de um catálogo de serviços. Identifica na literatura e propõe um modelo para a construção de catálogos de serviços de tecnologia da informação aplicável a organizações com o nível médio de maturidade das empresas brasileiras, baseado nas boas práticas preconizadas pelo Information Technology Infrastructure Library - ITIL, na sua versão 3.