| 摘要: |
| 采用RT-PCR和RACE-PCR技术克隆了大黄鱼(Larimichthys crocea)I 型干扰素基因全长cDNA序列及其基因组序列, 并利用荧光定量PCR技术研究了该基因在大黄鱼不同组织中的表达谱, 以及副溶血弧菌(Vibrio parahaemolyticus)、LPS、poly I:C刺激后大黄鱼脾脏、肝脏和头肾组织中该基因在转录水平的表达变化。结果表明, 大黄鱼I型IFN基因全长cDNA为878bp, 开放阅读框558bp, 编码185个氨基酸。推测的大黄鱼I型IFN氨基酸序列N端含有20个氨基酸的信号肽, 其后包含一个保守的IFabd结构域。大黄鱼I型IFN基因包含5个外显子, 4 个内含子, 在大多数组织和细胞中都有表达, 其中在血细胞中表达量最高, 肌肉中表达量最低。Poly I:C刺激可诱导大黄鱼I型干扰素基因在脾脏、头肾和肝脏中的表达量显著上调; LPS刺激可诱导大黄鱼I型干扰素基因在脾脏和肝脏中的表达量显著上调; 灭活的副溶血弧菌可诱导大黄鱼I型干扰素基因在头肾和肝脏中的表达量显著上调。 |
| 关键词: 大黄鱼 I型干扰素 结构特征 免疫反应 |
| DOI：10.11693/hyhz201302037037 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划(973)项目, 2011CB111604 号; 国家自然科学基金资助项目, 31101882 号 |
|
| MOLECULAR AND EXPRESSION CHARACTERIZATIONS OF TYPE I INTERFERON FROM LARGE YELLOW CROAKER LARIMICHTHYS CROCEA |
|
LI Chan1,2, YAO Cui-Luan1,2
|
|
1.Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture;2.Fisheries College,Jimei University
|
| Abstract: |
| The full length cDNA and genomic sequence of type I interferon (type I IFN) was cloned from large yellow croaker Larimichthys crocea by RT-PCR and RACE-PCR. The tissue expression and temporal expression profiles of LcIFN I gene after the stimulation with LPS, poly I:C and pathogenic bacteria Vibrio parahemolyticus were described. Our results showed that the full-length cDNA of LcIFN I was 878bp, and open reading frame (ORF) was 558bp, which encoded 185 amino acids. A signal peptide including 20 amino acid residues was at N-terminal, and an IFabd (Interferon alpha, beta and delta) domain was next to signal peptide. LcIFN I genomic sequence consisted of five exons and four introns. Quantitative real-time PCR analysis indicated a broad expression of LcIFN I in most detected tissues, with the most predominant expression in blood cells and the weakest expression in muscle. After injection with poly I:C, LcIFN I expression levels showed up-regulation in head-kidney, liver and spleen; After injection with LPS, LcIFN I expression levels showed up-regulation in liver and spleen; LcIFN I transcripts showed up-regulation in the liver and head-kidney after V. parahemolyticus injection. Our results suggested that type I IFN might play an important role in large yellow croaker anti-viral and bacterial immune response. |
| Key words: Larimichthys crocea Type I interferon characterization immune response |
