Selection of reliable reference genes for RT-qPCR studies in Octopus vulgaris paralarvae during development and immune-stimulation.

摘要: The common octopus, Octopus vulgaris is a new candidate species for aquaculture. However, rearing of octopus paralarvae hampered by high mortality and poor growth rates that impede its entire culture. study genes involved in the development immune response capability could help to understand key survival thus, complete life cycle. Quantitative real-time PCR (RT-qPCR) most frequently tool used quantify gene expression because specificity sensitivity. reliability RT-qPCR requires selection appropriate normalization whose must be stable across different experimental conditions study. Hence, aim present work evaluate stability six genes: β-actin (ACT), elongation factor 1-α (EF), ubiquitin (UBI), β-tubulin (TUB), glyceraldehyde 3-phosphate dehydrogenase (GADPH) ribosomal RNA 18 (18S) order select best reference gene. was analyzed using geNorm, NormFinder Bestkeeper, seven developmental stages (embryo, 0, 10, 15, 20, 30 34days) 20days after challenge with Vibrio lentus splendidus. results were validated measuring PGRP, stimuli-specific Our showed UBI, EF 18S as suitable during paralarvae, ACT bacterial infection. These provide basis further studies exploring molecular mechanism their innate defense.