Selection and validation of reference genes for functional studies in the Calliphoridae family

Autor(es)

Cardoso, Gisele Antoniazzi ^{[1, 2]} ; Matiolli, Cleverson Carlos ^{[1, 2]} ; Lima de Azeredo-Espin, Ana Maria ^{[1, 2]} ; Torres, Tatiana Teixeira ^{[3, 1]}

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas UNICAMP, CBMEG, Campinas, SP - Brazil

^[2] Univ Estadual Campinas UNICAMP, Dept Genet Evolucao & Bioagentes, Campinas, SP - Brazil

^[3] Univ Sao Paulo, Inst Biociencias, Dept Genet & Biol Evolut, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF INSECT SCIENCE; v. 14, JAN 2 2014.
• Citações Web of Science: 8

Resumo

The genera Cochliomyia and Chrysomya contain both obligate and saprophagous flies, which allows the comparison of different feeding habits between closely related species. Among the different strategies for comparing these habits is the use of qPCR to investigate the expression levels of candidate genes involved in feeding behavior. To ensure an accurate measure of the levels of gene expression, it is necessary to normalize the amount of the target gene with the amount of a reference gene having a stable expression across the compared species. Since there is no universal gene that can be used as a reference in functional studies, candidate genes for qPCR data normalization were selected and validated in three Calliphoridae (Diptera) species, Cochliomyia hominivorax Coquerel, Cochliomyia macellaria Fabricius, and Chrysomya albiceps Wiedemann. The expression stability of six genes (Actin, Gapdh, Rp49, Rps17, alpha-tubulin, and GstD1) was evaluated among species within the same life stage and between life stages within each species. The expression levels of Actin, Gapdh, and Rp49 were the most stable among the selected genes. These genes can be used as reliable reference genes for functional studies in Calliphoridae using similar experimental settings. (AU)

• Processo FAPESP: 08/58106-0 - Análise do perfil da expressão gênica em Cochiliomyia hominivorax através de sequenciamento massivo em paralelo
• Beneficiário: Tatiana Teixeira Torres
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/13463-3 - Expressão de genes relacionados ao comportamento alimentar na família Calliphoridae: uma análise em contexto filogenético
• Beneficiário: Gisele Antoniazzi Cardoso
• Modalidade de apoio: Bolsas no Brasil - Mestrado