tailieunhanh - Báo cáo khoa học: Molecular cloning and characterization of the crustacean hyperglycemic hormone cDNA from Litopenaeus schmitti Functional analysis by double-stranded RNA interference technique

The crustacean hyperglycemic hormone (CHH) plays an important role in the regulation of hemolymph glucose levels, but it is also involved in other functions such as growth, molting and reproduction. In the present study we describe the first CHH family gene isolated from the Atlantic Ocean shrimp Litopenaeus schmitti. | ỊFEBS Journal Molecular cloning and characterization of the crustacean hyperglycemic hormone cDNA from Litopenaeus schmitti Functional analysis by double-stranded RNA interference technique Juana M. Lugo1 Yuliet Morera2 Tania Rodríguez2 Alberto Huberman3 Laida Ramos2 and Mario P. Estrada1 1 Aquatic Biotechnology Department AnimalBiotechnology Division Center for Genetic Engineering and Biotechnology Havana Cuba 2 Marine Research Institute Havana University Cuba 3 NacionalNutrition Institute Salvador Zuriban Mexico DF Mexico Keywords cDNA amplification CHH double-stranded RNA Litopenaeus schmitti shrimp Correspondence M. P. Estrada Aquatic Biotechnology Project AnimalBiotechnology Division Center for Genetic Engineering and Biotechnology PO Box 6162 Havana 10600 Cuba Fax 53 7 2731779 Tel 53 7 2716022 Ext. 5154 E-mail Received 23 March 2006 revised 20 October 2006 accepted 25 October 2006 The crustacean hyperglycemic hormone CHH plays an important role in the regulation of hemolymph glucose levels but it is also involved in other functions such as growth molting and reproduction. In the present study we describe the first CHH family gene isolated from the Atlantic Ocean shrimp Litopenaeus schmitti. Sequence analysis of the amplified cDNA fragment revealed a high nucleotide sequence identity with other CHHs. Northern blot analysis showed that the isolated CHH mRNA from L. sch-mitti is present in the eyestalk but not in muscle or stomach. We also investigated the ability of dsRNA to inhibit the CHH function in shrimps in vivo. Injection of CHH dsRNA into the abdominal hemolymh sinuses resulted in undetectable CHH mRNA levels within 24 h and a corresponding decrease in hemolymph glucose levels suggesting that functional gene silencing had occurred. These findings are the first evidence that dsRNA technique is operative in adult shrimps in vivo. doi In crustaceans the X-organ-sinus gland complex in the eyestalk is

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