| 摘要: |
| 通过已构建的泥蚶血液cDNA文库, 克隆得到泥蚶Tg-HbIIA基因全长cDNA序列, 对其进行生物信息学、 组织表达及免疫相关分析。 结果表明, 泥蚶 Tg-HbIIA cDNA 全长 731bp, 包含 63bp 5′非编码区(5′-UTR), 246bp 3′非编码区(3′-UTR), 开放阅读框 450bp, 编码 150 个氨基酸; 其氨基酸序列与毛蚶(Scapharca kagoshimensis)和不等壳毛蚶(Scapharca inaequivalvis)的序列有较高的同源性, 分别达到 89%和 88%; 对其结构预测显示该蛋白具有血红蛋白功能域, 含有 10个潜在血红素结合位点。 qRT-PCR 检测结果显示: 泥蚶不同组织部位中, Tg-HbIIA mRNA 在血液表达量最大, 其次为外套膜和鳃, 表达量最低的是肝胰脏; 在副溶血弧菌(Vibrio parahaemolyticu)、脂多糖、肽聚糖刺激后, 泥蚶血液 Tg-HbIIA mRNA 表达量显著上调, 表明 Tg-HbIIA 在贝类抗菌免疫防御中可能起重要作用, 而 Tg-HbIIA 对不同致病因子免疫反应的灵敏度和表达时序存在一定差异。 |
| 关键词: 泥蚶, 血红蛋白, 菌刺激, mRNA 表达, 免疫反应 |
| DOI：10.11693/hyhz201201015015 |
| 分类号: |
| 基金项目:国家自然科学基金项目, 31001097 号; 浙江省自然科学基金项目, Y3100087 号; 国家贝类产业技术体系项目, CARS-48号。 |
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| CLONE AND ANALYSIS OF HEMOBLOBIN GENE (Tg-HbIIA) AND IMMUNE EXPRESSION RESEARCH IN TEGILLARCA GRANOSA |
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WANG Qing1,2, LIN Zhi-Hua1, BAO Yong-Bo1, HUO Li-Hui1,3, GU Hai-Long1,2
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1.College of Biological & Environmental Sciences, Zhejiang Wanli University;2.College of Fisheries and Life Sciences, Shanghai Ocean University;3.College of Life Sciences and Bioengineering, Ningbo University
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| Abstract: |
| The Tg-HbIIA cDNA was cloned and then the bioinformatics, expression and immune defense analysis of Tg-HbIIA cDNA were carried out. The full length of Tg-HbIIA cDNA was 731bp, which consists of an open reading frame of 450bp encoding a polypeptide of 150 amino acids, a 5'-UTR of 63bp and a 3'-UTR of 246bp; the deduced amino acid sequence of Tg-HbIIA shared 89% and 88% similarity with HbIIA from two species of the genus Scapharca respectively. The structure prediction of Tg-HbIIA showed the hemoglobin domain and the ten potential heme binding sites. The mRNA expression of Tg-HbIIA in haemocytes was detected highest in all the clam cells/tissues including haemocytes, adductor muscle, foot, hepatopancreas, gill and mantle. The Quantitative Real-time PCR showed that the mRNA transcript level of Tg-HbIIA, significantly up-regulated after Vibrio parahaemolyticus, Lipopolysaccharide and Peptidoglycan challenge which indicated that Tg-HbIIA was involved in the immune defense responses against bacterial infection and exposure to bacterial pathogenic factors. However, the temporal and level of expression were different in three challenge groups. |
| Key words: Tegillarca granosa, Hemoglobin, Bacteria challenge, mRNA expression, Immune response |
