O sequenciamento de nova geração tornou-se recentemente uma opção economicamente viável para a realização de aconselhamento genético, tendo proporcionado oportunidades sem precedentes para a pesquisa genômica personalizada, uma vez que o sequenciamento permite identificar a presença de variantes patogênicas. O teste genético que permite identificar as variantes patogênicas possui procedimentos laboratoriais como a coleta, preparação e sequenciamento do material e outros computacionais como o alinhamento, anotação e análise do exoma. A despeito do fato de que a grande quantidade de dados gerados no sequenciamento torna a atividade de análise relativamente complexa, esta tem sido realizada de forma manual por pesquisadores e profissionais da saúde, apesar de existir uma variedade de ferramentas computacionais disponíveis para auxiliar a análise e a interpretação de variações genéticas pontuais. Isto acontece porque a maioria destas ferramentas são generalistas, pouco amigáveis e não atendem às necessidades em pesquisa e atendimento clínico, não sendo utilizadas rotineiramente nesses ambientes. A análise manual, por sua vez, caracteriza-se por gerar resultados altamente dependentes tanto do conhecimento do pesquisador quanto da especificidade do procedimento realizado. O presente trabalho tem por objetivo desenvolver a modelagem do processo de análise de exoma, considerando a presença e a interação de diferentes papéis de usuário, num processo padronizado, porém flexível, juntamente com a implementação de uma ferramenta computacional amigável para a execução da análise baseada na modelagem proposta de forma organizada e otimizada, oferecendo suporte a atividades relacionadas à pesquisa e ao atendimento clínico. As análises processadas com a ferramenta também têm seus parâmetros e resultados armazenados e disponibilizados para uso em investigações futuras, possibilitando o compartilhamento do conhecimento para geração de novas descobertas. Nas avaliações realizadas para validação da proposta o modelo e a ferramenta mostram-se eficientes e apresentam boa usabilidade para a realização das análises / New generation sequencing has recently become an economically viable option for genetic counseling and has provided unprecedented opportunities for personalized genomic research since sequencing allows the identification of the presence of pathogenic variants. The genetic test that allows to identify the pathogenic variants has laboratory procedures like the collection, preparation and sequencing of the material and other computational as the alignment, annotation and analysis of the exoma. In spite of the fact that the large amount of data generated in the sequencing makes the analysis activity relatively complex, it has been performed manually by researchers and health professionals, although a variety of computational tools are available to aid analysis and the interpretation of specific genetic variations. This is because most of these tools are generalist, unfriendly and do not meet clinical research and care needs and are not routinely used in these settings. The manual analysis, in turn, is characterized by generating results highly dependent on both the knowledge of the researcher and the specificity of the procedure performed. The present work aims to develop the modeling of the exoma analysis process, considering the presence and interaction of different user roles, in a standardized but flexible process, together with the implementation of a user-friendly computational tool for performing the analysis based in the proposed modeling in an organized and optimized way, supporting activities related to research and clinical care. The analyzes processed with the tool also have their parameters and results stored and made available for use in future investigations, allowing the sharing of knowledge to generate new discoveries. In the evaluations carried out to validate the proposal, the model, and the tool are efficient and have good usability for carrying out the analysis

Le virus de la variole ovine est omniprésent dans les élevages de petits ruminants dans les pays d’Afrique du Nord et particulièrement en Tunisie malgré les campagnes de vaccination annuelles mises en place par les autorités vétérinaires du pays. L’optimisation de la souche vaccinale utilisée passe par le développement de vaccins dits de nouvelle génération tels que les vaccins sous unitaires utilisant des protéines reconnues pour induire une réponse humorale protectrice chez l’animal immunisé. Ceci pourrait être une alternative aux stratégies de lutte actuelles permettant de limiter la dissémination du virus en Tunisie. Peu de données existent sur les antigènes protecteurs spécifiques des virus du genre Capripoxvirus. Ce travail de thèse a ciblé, par homologie aux protéines du virus de la vaccine, quatre protéines du genre Capripoxvirus appartenant au virus de la dermatose nodulaire contagieuse potentiellement immuno-dominantes nommées LSDV60, LSDV117, LSDV122 etLSDV141 respectivement homologues des protéines L1, A27, A33 et B5. En premier lieu, une analyse structurale in silico a permis d’identifier les domaines essentiels de chaque protéine et de vérifier le taux de conservation de ces protéines parmi différents virus appartenant à la famille des poxvirus. Une analyse structurale approfondie mettant en évidence la structure primaire, secondaire et tertiaire de la protéine A27 a été réalisée. Suite à cette étude structurale, les protéines ont été produites dans deux systèmes d’expression différents ; le système eucaryote et le système baculovirus-cellules d’insectes afin de caractériser leur antigénicité vis-à-vis de sérums provenant d’animaux immunisés ou éprouvés.La reconnaissance des protéines d’intérêt en vecteur d’expression eucaryote n’a pas été concluante. En revanche, le système d’expression BEVS a permis la production de la protéine A27 (L1, A33 et B5 encours) avec succès sous forme soluble qui a été correctement reconnue par des sérums provenant de caprins naïfs challengés. La mise en évidence de formes trimériques et hexamériques confirment sonantigénicité. Une immunodétection des peptides correspondants à la protéine A27 synthétisés surmembranes (PepScan) combinée à une analyse in silico ont permis d'identifier des zones susceptibles de constituer des régions épitopiques reconnues situés majoritairement en partie N terminale de la protéine. / The sheep pox virus is omnipresent in small ruminant farms in North African countries andparticularly in Tunisia despite the annual vaccination campaigns set up by the Tunisian veterinaryauthorities. The optimization of the used vaccine strain involves the development of the so-called newgeneration vaccines such as subunit vaccines and this, using proteins recognized to induce a protectivehumoral response in the immunized animal. This could be considered as an alternative to currentcontrol strategies limiting virus spread in Tunisia. Few data exist on protective antigens specific toviruses in the genus Capripoxvirus. By homology to vaccinia virus proteins, this thesis work hastargeted four proteins in the genus Capripoxvirus belonging to the potentially immuno-dominantcontagious nodular dermatosis virus named LSDV60, LSDV117, LSDV122 and LSDV141respectively homologues of proteins L1, A27, A33 and B5. First, an in silico structural analysis hasallowed to identify the essential domains of each protein and to check the conservation rate of theseproteins among different viruses belonging to the poxvirus family. A thorough structural analysisidentifying the primary, secondary and tertiary structure of the A27 protein was conducted. Followingthis structural study, the proteins were produced in two different expression systems, namely theeukaryotic system and the baculovirus-insect cell system, in order to characterize their antigenicity tosera from immunized or proven animals. The recognition of the proteins of interest in the eukaryoticexpression vector has not been conclusive. On the other hand, the BEVS expression systemsuccessfully allowed the production of the A27 protein (L1, A33 and B5 in progress) in a solubleform, which was correctly recognized by sera from challenged naïve goats. Identifying trimeric andhexameric forms confirms its antigenicity. An immunodetection of the peptides corresponding toprotein A27 synthesized on membranes (PepScan) combined with an in silico analysis led to identifyzones capable of constituting recognized epitopic regions located predominantly in part N-terminal ofthe protein.