Resumo

ROMERO GERALDO, Reyna; HERNANDEZ SAAVEDRA, Norma; FIMBRES OLIVARRIA, Diana  e  GARCIA LAGUNAS, Norma. Validation of suitable reference genes for quantitative real-time PCR normalization in Crassostrea gigas spat stage during toxic dinoflagellates exposure. Biotecnia [online]. 2020, vol.22, n.2, pp.94-102.  Epub 07-Ago-2020. ISSN 1665-1456.  https://doi.org/10.18633/biotecnia.v22i2.1250.

The quantitative real-time polymerase chain reaction is a widely used method for gene expression analysis requiring carefully selected reference genes to ensure data validity. Regardless of several studies on gene expression bivalves and particularly Crassostrea gigas, has not been fully investigated regarding the evaluation of reference genes suitable for normalization of expression analysis. In this study, five candidate reference genes: actin, β tubulin, α subunit of elongation factor 1, glyceraldehyde-3-phosphate dehydrogenase and 28S ribosomal RNA were analyzed, to determine the most suitable reference genes, after of Crassostrea gigas spat were fed with Gymnodinium catenatum and Prorocentrum lima in mixed and compared to non-toxic diet Isochrysis galbana. The results showed that β-tub and ef-1α were the most stable genes for oysters feed with a mixed diet of P. lima and I. galbana. The gapdh and 28S rRNA were the most stable genes for oysters feed with G. catenatum and I. galbana. In addition, the selection of optimal reference genes during dinoflagellates exposure was verified by analyzing the expression level of trypsin and cytochrome c oxidase I target genes. Our study could be beneficial for future studies on gene expression in C. gigas.

Palavras-chave : Gene expression; reference gene; normalization; toxic dinoflagellates; Crassostrea gigas.

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