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.
Cloning of the crustacean hyperglycemic hormone gene promoter of Astacus leptodactylus
PEGORARO, SILVIA;GIULIANINI, PIERO GIULIO;EDOMI, PAOLO
2013-01-01
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.Pubblicazioni consigliate
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