Meeting Abstract
Crustaceans experience discontinuous growth because of the periodic shedding of their confining exoskeleton, a process known as molting or ecdysis. Because of periodic molting, much of crustacean physiology is cyclic, with many physiological and biochemical parameters varying during the molting cycle. House-keeping genes (HKGs) are commonly used as the internal control for quantitative real-time PCR (qPCR). In spite of the lack of assessment on the changeability of HKGs during crustacean molting cycle, several genes, including beta-actin, 18S rRNA and arginine kinase, have been used as the reference gene in qPCR quantification of expression of crustacean genes. The objective of this study was to determine which HKG is most stably transcribed during the crustacean molting cycle, using the blue crab, Callinectes sapidus, as the model crustacean. The expression stability of seven HKGs from the hepatopancreas, including beta-actin, glyceraldehyde 3-phosphate dehydrogenase, 28S rRNA, histone, ribosomal protein, arginine kinase and 18S rRNA, during the molting cycle of the blue crab, Callinectes sapidus, was assessed using the geNorm tool. The beta-actin gene gave rise to the lowest geNorm score, suggesting this gene is the most stably expressed, and therefore the best qPCR internal control of the seven HKGs tested.
