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2022, 42(5): 38-44. doi: 10.3969/j.issn.1673-9159.2022.05.005

墨吉明对虾高血糖激素基因FmCHH-I的分子鉴定和表达特征

广东海洋大学水产学院, 广东 湛江 524088

收稿日期:2021-07-11
修回日期:2021-07-11

基金项目:   广东省自然科学基金(2018A030310049);广东海洋大学南海青年学者专项 

关键词: 墨吉明对虾 , 高血糖激素 , CHH神经肽 , 基因表达

Molecular Identification and Expression of Hyperglycemic Hormone Gene in Fenneropenaeus merguiensis

College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China

Received Date:2021-07-11
Accepted Date:2021-07-11

Keywords: Fenneropenaeus merguiensis , crustacean hyperglycemic hormone , CHH neuropeptide , gene expression

【目的】研究墨吉明对虾(Fenneropenaeus merguiensis)高血糖激素基因FmCHH-I的分子特征和表达规律。【方法】根据转录组信息设计特异性引物,克隆FmCHH-I基因序列,并利用生物信息学手段预测其氨基酸和蛋白结构。采用实时荧光定量PCR方法分析该基因在不同组织中和卵巢发育不同时期的表达情况,以及切除单侧眼柄对其表达的影响。【结果与结论】成功克隆墨吉明对虾高血糖激素基因FmCHH-I的序列。该基因cDNA全长599 bp,开放阅读框(ORF)长为369 bp,编码123个氨基酸。蛋白序列的1~20位包含一个信号肽,随后有一段由26个氨基酸组成的前导肽,末端为成熟肽,包含74 个氨基酸以及6 个半胱氨酸残基。FmCHH-I 基因与斑节对虾(Penaeus monodonCHH-I 基因同源性最高(91.53%),与欧洲龙虾(Homarus gammarusCHH-I 基因同源性最低(43.86%)。系统发育分析表明,FmCHH-I 与斑节对虾聚在一起,与对虾科物种聚为一支。FmCHH-I 在眼柄中的表达量最高,在肝胰腺、肠道中也有表达。FmCHH-I 在性腺未发育期和性腺成熟期雌性对虾眼柄中的表达量无显著差异(P > 0.05),但切除眼柄后,卵巢中FmCHH-I的表达量显著上调(P < 0.01)。

【Objective】 To study the molecular characteristics and expression pattern of the hyperglycemic hormone gene FmCHH-I in the shrimp Fenneropenaeus merguiensis.【Methods】Specific primers were designed based on transcriptome information for cloning the FmCHH-I gene sequence.Bioinformatic prediction was utilized to analyze its amino acid and protein structure.Realtime fluorescence quantitative PCR was used to analyze its expression profiles in different tissues and at various stages of ovary development.Its expression change following unilateral eyestalk removal was also investigated.【Results and Conclusions】 The sequence of the hyperglycemic hormone gene FmCHH-I was successfully cloned.The full-length cDNA was 599 bp, and the open reading frame(ORF) was 369 bp long, encoding 123 amino acids.The 1 -20 positions of the protein sequence contained a signal peptide, followed by a leading peptide composed of 26 amino acids and a mature peptide at the end, including 74 amino acids and 6 cysteine residues.The FmCHH-I gene had the highest homology with Penaeus monodon's CHH-I gene (91.53%), and the lowest homology with Homarus gammarus's CHH-I gene(43.86%).Phylogenetic analysis showed that FmCHH-I was closely related to its homologue from the giant tiger shrimp, and formed a cluster with homologous genes from Penaeidae species.There was no significant difference in the expression of FmCHH-I in the eyestalk of female shrimp between the undeveloped stage and the ovarian maturation stage (P > 0.05), but its expression was significantly induced in the ovary after removal of the eyestalk(P < 0.01).

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墨吉明对虾高血糖激素基因FmCHH-I的分子鉴定和表达特征