锦鲤Hepcidin基因的克隆及其组织特异性表达分析（英文）

发布时间：2018-07-22 19:40

【摘要】：【目的】克隆锦鲤Hepcidin全长c DNA序列(k-hepc),并获得此基因在鱼体内的表达模式。【方法】利用RT-PCR和RACE PCR的方法,从锦鲤肝脏中克隆锦鲤Hepcidin的全长c DNA,进行序列测定和分析;锦鲤经肌肉注射维氏气单胞菌0、4、8、12、24和48 h后,分别取其肝、脾、肾、肠、脑、心、肌肉和鳃组织,采用实时荧光定量PCR的方法,以β-actin为内参基因,检测k-hepc基因的表达量。【结果】锦鲤抗菌肽(Gen Bank登录号KC795559)全长755 bp,编码序列276 bp,编码91个氨基酸,包括信号肽、原肽和成熟肽,成熟肽C端含有8个半胱氨酸,可形成4个分子内二硫键。与已报道的普通鲤鱼Hepcidin氨基酸序列的一致性为93%,与其他鱼类Hepcidin氨基酸序列的一致性为29%-93%。在本研究所检测的正常锦鲤的组织中,k-hepc均有表达,其中在肝组织中表达量最高,鳃组织中表达量最低。经维氏气单胞菌感染后,k-hepc在肝和心组织中的表达量明显增加,在其余组织中变化不显著。【结论】k-hepc编码的蛋白是Hepcidin家族的成员之一。锦鲤Hepcidin的表达主要受内在调节因素影响。
[Abstract]:[objective] to clone the full-length c-DNA sequence (k-hepc) of koi carp (Cyprinus Carpio) and obtain its expression pattern in fish. [methods] the full-length c-DNA of Koi carp Hepcidin was cloned from the liver of koi carp by RT-PCR and race PCR, and the sequence was sequenced and analyzed. The liver, spleen, kidney, intestine, brain, heart, muscle and Gill tissues of koi were injected intramuscularly with Aeromonas Vickers 04 (Amonomonas Vickers) strain 04, 1212, 24 and 48 h later, and 尾 -actin was used as internal reference gene by real-time fluorescence quantitative PCR (PCR), and 尾 -actin (尾 -actin) was used as internal reference gene. [results] the total length of antimicrobial peptide of koi (GenBank accession number KC795559) was 755 BP, encoding 276bp. encoding 91 amino acids, including signal peptide, propeptide and mature peptide, and the C-terminal of mature peptide contained 8 cysteine. Four intramolecular disulfide bonds can be formed. The consistency of the amino acid sequence with the reported common carp was 933 and that with the other fish was 29- 93.The consistency of the amino acid sequence with the reported common carp was 933. In this study, the expression of khepc was found in the normal koi tissues, the highest in the liver and the lowest in the Gill. After infection with Aeromonas Vickers, the expression of k-hepc in liver and heart was significantly increased, but not in other tissues. [conclusion] the protein encoded by k-hepc is one of the members of the hepin family. The expression of koi hepcidin was mainly affected by internal regulatory factors.
【作者单位】：北京市水产科学研究所渔业生物技术北京市重点实验室;中国水产科学研究院生物技术研究中心;
【基金】：The Ornamental Fish Innovation Consortium of Beijng(No.BAIC03-2017) Natural Science Foundation of Beijng(No.6152006)~~
【分类号】：S917.4

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