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Doctoral Thesis
DOI
https://doi.org/10.11606/T.17.2020.tde-08012020-104240
Document
Author
Full name
Henrique Oliveira Frank
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2019
Supervisor
Committee
Monesi, Nadia (President)
Larson, Maria Luisa Paçó
Silva, Luis Lamberti Pinto da
Simoes, Zila Luz Paulino
Siviero, Fábio
Title in Portuguese
Genes responsivos ao hormônio ecdisona em Bradysia hygida (Diptera, Sciaridae)
Keywords in Portuguese
Bradysia hygida
Ecdisona
Genes de resposta primária
Sciaridae
Abstract in Portuguese
Este trabalho visa a identificação de genes responsivos a via de transdução de sinal do hormônio ecdisona no sciarídeo Bradysia hygida. Os genes EcR, USP, E75, E74 e Broad-complex foram os selecionados para dar início a caracterização desta via em B. hygida. Inicialmente, através de abordagens in silico foi possível elaborar a estrutura gênica e suas respectivas isoformas. Para tal, foram utilizados como ponto de partida sequências brutas de 2 transcriptomas depositados do também sciarídeo B. odoriphaga, além de sequências de outros insetos dípteros como Drosophila melanogaster e Aedes aegypti, porém os dados finais foram obtidos utilizando somente por buscas no genoma de B. hygida, com o transcriptoma de embriões de B. hygida. Através de buscas no genoma montado de B. hygida foi possível identificar em diferentes segmentos genômicos os diferentes genes alvo. Foram identificadas 3 isoformas, que codificam 2 diferentes polipeptídeos, para o gene USP. Também foram identificadas quatro diferentes isoformas para o gene EcR, nomeadas A1, A2, A3 e B, visto que as 3 pertencentes a isoforma EcRA diferiam somente em suas regiões 5' UTR. Para a identificação dos genes E74, E75 e Broad-Complex, também foram utilizadas sequências obtidas no transcriptoma de embriões de B. hygida. Obteve-se um total de 2 isoformas de E74 (A e B), 5 possíveis isoformas para E75 (A, B, C, D, E) e, também 5 possíveis (A, B, C, D, E) para o gene Broad-Complex em B. hygida. Com isso, pretende-se estender a caracterização da rede reguladora de expressão gênica induzida pelo hormônio ecdisona e futuramente caracterizar a expressão destes genes em diferentes tecidos. No momento os dados in silico permitem afirmar que a via descrita em outros insetos parece estar conservada em B. hygida, o que nos permitirá produzir dados inéditos a respeito deste díptero basal e ampliar a caracterização de genes iniciais responsivos à ecdisona, durante a metamorfose.
Title in English
The ecdysone hormone responsive genes in Bradysia hygida (Diptera, Sciaridae)
Keywords in English
Bradysia hygida
Ecdysone
Initial genes
Sciaridae
Abstract in English
This work aims to identify responsive genes from the transduction signal pathway of the ecdysone hormone in the sciarid Bradysia hygida. The genes EcR, USP, E75, E74 e Broad-complex were selected to begin the characterization of this pathwa in B. hygida. Initially, through in silico approaches it was possible to elaborate the gene structure and also there respective isoforms. Doing so, it was used as a starting point 2 groups of raw sequences from transcriptomes from the also sciarid B. odoriphaga, and also sequences from other dipterian insects as Drosophila melanogaster and Aedes aegypti, however the final data were obtained only by searching the genome of B. hygida, with the transcriptome of embryos of B. hygida. Searching the assembled genome from B. hygida it was possible to identify on diferente genomic segments the target genes. Three isoforms, which encode 2 different polypeptides, were identified for the USP gene. We also identified four different isoforms for the EcR gene, named A1, A2, A3 and B, since the 3 belonging to the EcRA isoform differed only in their 5 'UTR regions. For the identification of E74, E75 and Broad-Complex genes, sequences obtained in the transcriptome of B. hygida embryos were also used. A total of 2 isoforms of E74 (A and B), 5 possible isoforms for E75 (A, B, C, D, E) and also 5 (A, B, C, D, E) isoforms were obtained for Broad-Complex gene in B. hygida. With this, we intend to extend the characterization of the regulating network of gene expression induced by the hormone ecdysone and to characterize the expression of these genes in different tissues in the future. At the moment the in silico data allow us to affirm that the pathway described in other insects seems to be conserved in B. hygida, which will allow us to produce unprecedented data about this basal dipteran and to extend the characterization of initial genes responsive to ecdysone, during metamorphosis.
 
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Publishing Date
2020-04-29
 
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