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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2009.tde-01022010-092416
Document
Author
Full name
Jeniffer Novaes Gonçalves Dias
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Madeira, Alda Maria Backx Noronha (President)
Carboni, Beatriz Simonsen Stolf
Ho, Paulo Lee
Menck, Carlos Frederico Martins
Ojopi, Elida Paula Benquique
Title in Portuguese
Análise da expressão diferencial entre merozoítos e esporozoítos de Eimeria tenella empregando a técnica de LongSAGE.
Keywords in Portuguese
Eimeria tenella
Coccidiose
Esporozoítos
Expressão Gênica
Merozoítos
Abstract in Portuguese
Eimeria tenella é umas das principais espécies que causam a coccidiose aviária. Para se estudar o perfil de expressão gênico quantitativo em estágios infectantes bibliotecas de LongSAGE foram geradas a partir de merozoítos e esporozoítos. Mais de 35.000 tags foram obtidas, das quais, 9.516 eram únicas. Para a identificação e anotação de genes diferencialmente expressos, as tags foram extraídas, contadas e analisadas estatisticamente por um pacote desenvolvido pelo nosso grupo, SAGE Analysis. Um total de 197 seqüências foram reconstruídas e anotadas automaticamente. Foi observado uma expressão estágio-específica e perfil transcricional distinto entre os estágios. Em merozoítos, foram encontradas proteínas envolvidas na tradução e manutenção da conformação protéica e em esporozoítos, os resultados positivos foram relacionados à cromatina, transporte e atividade catalítica. Para validação da técnica, a expressão diferencial de um pequeno conjunto de genes foi quantificada por RT-qPCR. Os resultados demonstraram uma boa correlação entre estas duas plataformas.
Title in English
Differential expression analysis between merozoites and sporozoites of Eimeria tenella using LongSage.
Keywords in English
Eimeria tenella
Coccidiosis
Gene Expression
Merozoites
Sporozoites
Abstract in English
Eimeria tenella is one of the most important causing agents of poultry coccidiosis. To study the quantitative gene expression profile in zoite stages of LongSage libraries were generated from merozoites and sporozoites. More than 35.000 tags were obtained, whose 9.516 were unique. For identification and annotation of differential expressed genes, tags were extracted, counted and submitted to statistical analysis by Sage Analysis, software developed by our group. A total of 197 tags were reconstructed and automatic annotated. Stage-specific expression genes and distinct transcriptional profile were observed between these stages. In merozoites the results were related to protein translation and folding, and in sporozoites the proteins were involved to chromatin structure, transport and catalytic activity. To LongSAGE validation, differential expression was quantified using RT-qPCR to a small group of genes. Good correlation was observed between these platforms.
 
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Publishing Date
2010-03-01
 
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