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本文首次采用RACE技术获得口虾蛄(Oratosquilla oratoria)眼柄部高血糖激素(crustacean hyperglycemic hormone,CHH)基因cDNA全长序列,共2 421bp,5′UTR长度为180bp,3′UTR为1 821bp,开放阅读框(ORF)长420bp,编码139个氨基酸。预测蛋白二级结构α-螺旋23.74%,延伸链占23.02%,无规则卷曲占53.24%,预测分子量为15.47kD,等电点(pI)为7.049。CHH成熟肽包含6个CHH家族中位置保守的Cys残基,C端为GK,推断为ES型CHH基因。聚类分析表明:口虾蛄CHH和十足目CHH聚为同一个分支的两亚群,具有较近的亲缘关系。荧光定量PCR分析结果表明,CHH基因在肠中表达量最高,在眼柄、触角中有较高的表达量,在肌肉中表达量相对较低,在卵巢中表达最低,而在精巢中不表达。
In this paper, the full-length cDNA of CHH gene in eyeballs of Oratosquilla oratoria was obtained for the first time using RACE technique, with a total length of 421bp, length of 5’UTR of 180bp and length of 3’UTR of 1821bp, The open reading frame (ORF) is 420 bp long and encodes 139 amino acids. The predicted secondary structure of the protein was 23.74% in α-helix, 23.02% in extension chain and 53.24% in random coil. The predicted molecular weight was 15.47kD and the isoelectric point (pI) was 7.049. The CHH mature peptide contains six conserved Cys residues in the CHH family, the C-terminal GK, and the ESH CHH gene. Cluster analysis showed that both CHH and CHH of shrimp 虾 蛄 clustered into two sub-populations of the same branch, which had close genetic relationship. The result of real-time quantitative PCR showed that CHH gene had the highest expression in the intestine, higher expression in the eyelid and antennae, lower expression in muscle, lowest expression in ovary and no expression in testis .