Cloning of the crustacean hyperglycemic hormone gene promoter of Astacus leptodactylus

Abstract

The crustacean Hyperglycemic Hormone (cHH) belongs to a family of neuropeptides synthesized and released by the X-organ sinus gland complex, which is located in the eyestalks. The cHH is a pleiotropic hormone regulating many physiological processes like glucose metabolism, molting, and reproduction. We cloned a 176 bp fragment from the transcription starting site of the cHH gene of Astacus leptodactylus Eschscholtz, 1823. The identified sequence contains several putative core transcription factor binding sites including a TATA box, a CCAAT box, a GC box, as well as binding sites for a hypoxia responsive element, estrogen receptor, and ecdysone receptor. To assess promoter functionality the promoter sequence was ligated to an expression vector bearing a luciferase reporter gene, and promoter activity was recorded using a luciferase expression assay, which showed that the cloned promoter was functional, being able to drive the expression of the reporter gene.