摘要: |
白细胞介素7受体α链(IL-7Rα,CD127)是一个重要的多效性细胞因子受体.本研究克隆得到大黄鱼(Larimichthys crocea)的IL-7Rα(LycIL-7-Rα),其开放阅读框全长1 242 bp,编码413个氨基酸.通过氨基酸序列分析和多序列比对发现,LycIL-7Rα多肽具有I型细胞因子受体的典型特征,包括4个保守的半胱氨酸、1个信号肽、1个单次跨膜结构域、1个胞外的fibronectin type III(FN III-like)结构域、1个Trp-Ser-X-Trp-Ser (WSXWS)基序以及胞内Box1结构域.系统进化分析表明,LycIL-7Rα与其他鱼类的IL-7Rα聚在一起,与鲈鱼(Lates calcarifer)IL-7Rα的亲缘关系最近.Real-time PCR结果表明,LycIL-7-Rα在健康大黄鱼中为组成型表达,在脾脏中的表达量最高,在心脏中的表达量最低.大黄鱼经聚肌苷酸胞苷酸[Poly(I:C)]或溶藻弧菌(Vibrio alginolyticus)刺激后,其脾脏中LycIL-7Rα的mRNA转录水平均显著上调.这些结果表明 LycIL-7Rα可能在大黄鱼免疫反应中发挥重要作用. |
关键词: 海洋生物学 大黄鱼 白细胞介素7受体α链 分子克隆 实时定量PCR 表达分析 |
DOI:10.3969/J.ISSN.2095-4972.2018.02.016 |
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基金项目:国家自然科学基金资助项目(31602194);海洋经济创新发展区域示范项目资助项目(2014FJPT10) |
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Cloning and expression analysis of Larimichthys crocea IL-7-Rα gene |
LIU Ying-di,ZHAO Qun-fen,YANG Jun-peng,CHEN Xin-hua,AO Jing-qun |
(School of Marine Sciences, Ningbo University, Ningbo 315211, China;Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, SOA, Xiamen 361005, China) |
Abstract: |
The IL-7 receptor alpha chain (IL-7Rα,CD127) is an important pleiotropic receptor in immunology. In this paper, we cloned an IL-7Rα homologue from Larimichthys crocea and named it as LycIL-7-Rαv. The deduced open reading frame (ORF) of LycIL-7Rα is 1 242 base pair (bp) long, encoding a protein consisting of 413 amino acids (aa). Amino acid sequence alignment revealed that LycIL-7Rα protein exhibits typical domain architecture of type I cytokine receptors, including 4 conserved cysteine residues, a signal peptide, an extracellular FN III-like domain, a Trp-Ser-X-Trp-Ser (WSXWS) motif, a transmembrane region and an intracellular Box1 domain. Phylogenetic analysis shows that LycIL-7Rα fell into a clade formed by all fish IL-7Rα and was closely related to IL-7Rα of barramundi perch (Lates calcarifer). Real-time PCR results shows that LycIL-7RαmRNA was constitutively expressed in all tissues tested, with highest expression level in spleen and lowest in heart. Moreover, the transcription levels of LycIL-7Rα mRNA are significantly upregulated in spleen after stimulated by polyinosinic acid cytidine [Poly (I:C)] or Vibrio alginolyticus. These results indicate that LycIL-7Rα may play roles in immune responses in the large yellow croaker. |
Key words: marine biology Larimichthys crocea interleukin-7 receptor alpha chain molecular cloning real-time PCR expression analysis |