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Genes Diferencialmente Expressos Em Paracoccidioides Brasiliensis

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Genes Diferencialmente Expressos Em Paracoccidioides Brasiliensis UNIVERSIDADEDE BRASÍLIA FACULDADEDE MEDICINA PROGRAMA DEPÓS-GRADUAÇÃO EM PATOLOGIA MOLECULAR Genes diferencialmente expressos em Paracoccidioides brasiliensis TESE DE DOUTORADO Candidato: CLAYTON LUIZBORGES Orientadora:Dra. CÉLIA MARIA DE ALMEIDA SOARES Novembro - 2008 Livros Grátis http://www.livrosgratis.com.br Milhares de livros grátis para download. UNIVERSIDADE DE BRASÍLIA FACULDADE DE MEDICINA PÓS–GRADUAÇÃO EM PATOLGIA MOLECULAR Genes diferencialmente expressos em Paracoccidioides brasiliensis Candidato: Clayton Luiz Borrges Orientadora: Profa. Dra. Célia Maria de Almeida Soares Tese apresentada ao Programa de Pós-Graduação em Patoloogia Molecular como requisito para obtenção do grau de Doutor em Patologia Molecular Brasília-DF-Brasil -2008- TRABALHO REALIZADO NO LABORATÓRIO DE BIOLOGIA MOLECULAR, DO DEPARTAMENTO DE BIOQUÍMICA E BIOLOGIA MOLECULAR, DO INSTITUTO DE CIÊNCIAS BIOLÓGICAS DA UNIVERSIDADE FEDERAL DE GOIÁS Apoio Financeiro: CNPq/ FINEP/SECTEC-GO Brasília-DF-Brasil -2008- II UNIVERSIDADE DE BRASÍLIA FACULDADE DE MEDICINA PÓS–GRADUAÇÃO EM PATOLGIA MOLECULAR BANCA EXAMINADORA TITULARES: Profa. Dra. Célia Maria de Almeida Soares Instituto de Ciências Biológicas, Universidade Federal de Goiás Profa. Dra. Patrícia Silva Cisalpino Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais Prof. Dr. Cézar Martins de Sá Instituto de Ciências Biológicas, Universidade de Brasília Profa. Dra. Anamélia Lorenzetti Bocca Faculdade de Medicina, Universidade de Brasília Profa. Dra. Sílvia Maria Salem-Izacc Instituto de Ciências Biológicas, Universidade Federal de Goiás SUPLENTE Prof. Dr. Bergmann Morais Ribeiro Instituto de Ciências Biológicas, Universidade de Brasília III ʺSuperação é poder fazer acontecer com as ferramentas que temos em mãos, Superação é trabalhar da melhor forma possível independentemente do que pensem ou falem. Superação é irmos além do que os outros acham que somos capazes!!!ʺ (Autor desconhecido) ʺVencer a si próprio é a maior de todas as vitóriasʺ Platão Para ser feliz basta ter amigos, família e orgulho do que se faz... ...sou feliz! Claytim IV Amar é sentir na felicidade do outro a própria felicidade. Gottfried Wilhelm von Leibnitz Dedico este trabalho à minha família: Ao meu pai, minha referência de honestidade, amor e dedicação; e que mesmo ausente sempre esteve conosco; À minha mãe, que sem meu pai e grávida do caçula da família, não mediu esforços para nos fornecer o básico e o mais importante: o amor. À minha esposa, você é o maior exemplo de que o amor supera tudo. Você me entende, me auxilia, fica comigo e me deixa quando preciso. Sem você eu não teria chegado aqui. Nossa união é para sempre! Amo você! À minha filha, que me chamou tantas vezes para brincar, jogar vídeo‐game e passear... diante do meu não sorriu e me entendeu. Papai te ama!! Aos meus tios (Eurípedes e Desnaídes), meus “segundos pais”; vocês sempre foram uma referência para mim. Aos meus padrinhos que “andaram sumidos” e retornaram para nosso convívio familiar. Que bom! Aos meus irmãos, Alexandre e Leonardo. Alexandre, você é meu exemplo de serenidade e paciência; e você Leo, exemplo de inteligência e dedicação. Amo muito todos vocês !!! V AGRADECIMENTOS A Deus, pelas infinitas bênçãos, pela força e energia presentes todos os dias e por ter direcionado todos os meus caminhos; Há 15 anos conheci uma pessoa... eu era “office boy”. Ela viajou pro Canadá (eu ajudei a encaminhar a reserva da passagem dela)... acho que para participar de um Congresso... quando ela voltou, me trouxe um chaveiro de Vancouver; que honra!!! Depois meu chefe, o Dr. Mário Okuda, me disse: “Essa professora é umas das poucas cujo comportamento deve ser copiado aqui”... Isso me levou a querer ser assim também... Anos depois estou aqui..... Professora Drª Célia Maria de Almeida Soares, minha orientadora... você é um exemplo a ser seguido; a grande responsável pela minha formação. Com você aprendi não só a “pipetar”... aprendi que tem que se dar oportunidades para quem quer e para quem merece... aprendi que só com trabalho e muito esforço podemos ser competitivos e superar nossas dificuldades... aprendi que a cobrança é proporcional à confiança... aprendi que o bem público deve ser usado com seriedade e honestidade... Célia, sinto-me honrado de participar do seu grupo. Obrigado por tudo!!!! Nem sei como falar de uma pessoa que admiro tanto.... Meu melhor amigo, um irmão, um companheiro.... não sabe? Pergunta pra ele!!! Trabalhamos muito juntos cara... tantas noites... tantos finais de semana no laboratório... quantos congressos juntos... ainda vamos trabalhar muito juntos meu irmão. Obrigado por ser o que você é... por ter me ajudado e ter dado a oportunidade de convivermos para que pudesse aprender um pouco com você. Tenho orgulho de ser seu amigo Alex Betão! Às minhas filhinhas do coração... Elisa e Mirelle, com vocês aprendi mais do que ensinei; À Juju (minha irmãzinha) e a Fafah (um doce de pessoa). Juju, você fez tanto por mim. Muito obrigado pela convivência!!! Ao Rodriguinho, um cara muito especial, menino mais com muito potencial. Demos muitas gargalhadas juntos; VI Meus agradecimentos à Moc, minha colega na graduação, minha amiga que ajudou tanto na minha formação; Aos meus amigos Luiz (Dindin – o cara! Detalhe!) e Wesley (Tchatchá-pesquisador), vocês tornaram os momentos de trabalho muito melhores; À Dani e a Renata, pessoas muito importantes na minha formação. Tivemos bons momentos na bancada e fora dela; Aos queridos companheiros do LBM: Tereza Cristina, Nathalie, Ronney, Sara, Daciene, Milce, Karinne, Bernadette, Patrícia Lima, Mariana, Nadya, Bruno, Kelly, Ivian, Yuri, Gabi, Moniquinha, Leandro, Daiane, Hellen, Ademar, Heiko, Patrícia Kott, Rogério, Renata, Regilda, Aline, Kesser, Patrícia Zambuzzi, Amanda, Raquel, Neto, Christielly, Hérika, João Guilherme e Lidiane. Muitos alunos, muitas diferenças e muito aprendizado! Aos membros da Banca que aceitaram nosso convite para contribuir com nosso trabalho; À todos os professores do programa de Pós-Graduação em Patologia Molecular-UnB, os quais foram de suma importância na minha formação; Aos funcionários da Secretaria da Pós – Graduação em Patologia Molecular, Hérica, Alessandro e Jaqueline, sempre atenciosos, eficientes e auxiliando todos do grupo de Goiânia. À Profa. Gláucia Maria Gleibe de Oliveira, do DPP-UnB, pela presteza cordialidade; À todos os meus colegas, alunos de Mestrado e Doutorado: crescemos juntos nos trabalhos, seminários e aulas; À Dona Zildete (in memoriam) e à Dona Dora pelo auxílio prestado aos alunos; À todos os meus familiares, amigos e colegas e as pessoas que, de alguma maneira participaram desta jornada; À todos, MUIITO OBRIIGADO! VII SUMÁRIO Página Lista de Abreviaturas .............................................................................................. X Resumo ................................................................................................................... XIII Abstract................................................................................................................... XIV CAPÍTULO I I. Introdução I.1. Aspectos Gerais.................................................................................... 15 I.2. Paracoccidioidomicose.......................................................................... 15 I.3. Classificação Taxonômica..................................................................... 16 I.4. Biologia de P. brasiliensis..................................................................... 18 I.5. Dimorfismo........................................................................................... 19 I.6. Projetos Transcritomas de P. brasiliensis............................................. 22 I.7. Genes diferencialmente expressos no processo infeccioso................. 25 I.8. RDA (Análise de Diferença Representacional) no estudo de genes 27 diferencialmente expressos................................................................................... I.9. Formamidase: gene diferencialmente expresso em P. brasiliensis..... 30 II. Justificativas................................................................................................... 32 III. Objetivo Geral.............................................................................................. 33 III.1. Objetivos Específicos........................................................................ 33 CAPÍTULO II Artigos – Formamidase de P. brasiliensis ................................................. 34 Discussão.................................................................................................... 57 Conclusões.................................................................................................. 61 CAPÍTULO III Artigo – Análises transcricionais no estudo de genes diferencialmente 63 expressos em P. brasiliensis.................................................................................. Discussão.................................................................................................... 80 Conclusões.................................................................................................. 83 VIII CAPÍTULO IV Artigo com autoria compartilhada............................................................. 84 CAPÍTULO V Artigos publicados em colaboração no período de realização do Doutorado 85 CAPÍTULO VI IV. Perspectivas.......................................................................................... 86 V. Referências Bibliográficas....................................................................
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