中国水产科学2016，Vol.23Issue(2)：307-315,9.DOI:10.3724/SP.J.1118.2016.15149
三疣梭子蟹核受体基因HR38的克隆及其在蜕皮中的表达分析
Cloning and expression analysis in molting cycle of nuclear receptor HR38 gene inPortunus trituberculatus
摘要
Abstract
The nuclear receptor superfamily contains a large number of transcription factors that mediate many physio-logical process, such as cell differentiation, growth and development. Family members have a conserved functional domain that includes a DNA-binding domain and a ligand-binding domain. HR38, a member of the nuclear receptor superfamily, is an orphan receptor for which no ligand is known. HR38 is involved in an unusual ecdysteroid signaling pathway in Drosophila.Portunus trituberculatus is a major Chinese aquaculture species and is also an important Chi-nese export. The crustacean molting cycle is a physiologically important growth process. To investigate its function in the molting cycle ofPortunus trituberculatus, cDNA encoding nuclear receptor HR38 ofPortunus trituberculatus was cloned by rapid amplification of cDNA ends (RACE) and namedPHR38. The full-lengthPTHR38 cDNA is 2950 bp, including a 101-bp 5′ untranslated region (UTR) and a 551-bp 3′ UTR. The 2298-bp open reading frame (ORF) encodes a 765-amino acid polypeptide. Bioinformatic analysis revealed that PTHR38 is anunstable protein and has no trans-membrane domains. Homology analysis showed thatPTHR38 ofPortunus trituberculatus has the highest homology to the HR38 gene ofDaphnia magna. Six crabs ateach main molting stage (inter-, pre- and post-molting) were selected by morphological feature observation. Eyestalks were excised, the crabs were depigmented, their exoskeletons were re-moved, and different tissues were sampled before being stored in Trizol Reagent for RNA extraction. Quantitative real-time PCR showed different expression patterns ofPTHR38at each stage of the molting cycle, which suggested that PTHR38 functions in the molting cycle ofPortunus trituberculatus. It also showed a similar expression tendency to ecdysone in hemocytes and thus might be associated with ecdysone. In muscle and the hepatopancreas,PTHR38was upregulated in the post-molting stage. Previous studies showed that HR38 participates in the metabolism of carbohy-drates, which suggested thatPTHR38might function in energy metabolism processes. After eyestalk ablation, the ex-pression ofPTHR38was upregulated, which also suggested a connection betweenPTHR38 and ecdysone.关键词
三疣梭子蟹/核受体/HR38/基因克隆/基因表达Key words
Portunus trituberculatus/nuclear receptor/HR38/gene cloning/gene expression分类
农业科技引用本文复制引用
张龙涛,吕建建,高保全,刘萍,付萍..三疣梭子蟹核受体基因HR38的克隆及其在蜕皮中的表达分析[J].中国水产科学,2016,23(2):307-315,9.基金项目
国家自然科学基金资助项目(41306177 （）
41576147) （）
