| 摘要: |
| 经转录组测序后筛选并克隆得到青蛤(Cyclina sinensis)髓样分化因子88(myeloid differenttiationfactor 88, MyD88)的cDNA 序列。在鳗弧菌(Vibrio anguillarum)胁迫下, 采用荧光定量PCR 法分析了MyD88 基因在青蛤体内的表达过程。结果显示, 青蛤MyD88 基因的开放阅读框为1521bp, 编码506 个氨基酸, 分子量约为57.14kDa, 氨基酸N 段存在DEATH 结构域, C 段存在TIR 结构域(Toll/Interleukin-1 receptor domain)。MyD88 基因在青蛤血淋巴、肝脏、外套膜、鳃和闭壳肌等组织中普遍表达, 但在血淋巴中表达量最高, 与其它组织出现显著性差异(P<0.05)。通过检测鳗弧菌刺激下MyD88 基因在青蛤血淋巴中的表达值, 发现MyD88 基因在24h 开始升高, 48h 达到最大值, 约为对照组的10 倍, 实验组与对照组及空白组均出现了极显著性差异(P<0.01); 研究结果表明, 该基因在软体动物的免疫应答反应中对革兰氏阴性菌有识别作用。 |
| 关键词: 青蛤 鳗弧菌 转录组文库 MyD88 荧光定量PCR |
| DOI：10.11693/hyhz20141000290 |
| 分类号: |
| 基金项目:天津市科委应用基础与前沿技术重点项目资助,12JCZDJC22800号。 |
附件 |
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| EFFECT OF VIBRIO ANGUILLARUM ON GENE EXPRESSION OF MyD88 IN CYCLINA SINENSIS |
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GAO Jing, REN Yi-Peng, PAN Bao-Ping, YAN Chun-Cai
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College of Life Sciences, Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University, Tianjin 300387, China
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| Abstract: |
| A gene of Cyclina sinensis myeloid differentiation factor 88 (CsMyD88) was cloned and analyzed with transcriptome library for studying the function in innate immune response. The expression of CsMyD88 was detected by real-time quantitative PCR. Results show that the CsMyD88 was express to the highest level in hemolymph significantly different from those in liver, adductor muscle, gills, and mantle (P<0.05). The expression of CsMyD88 in hemolymph injected with Vibrio anguillarum increased in 24h and peaked in 48h, which is significantly different (P<0.01) from that of the control. Therefore, Vibrio caused tissue injury at the first stage of infection, to which CsMyD88 responded and expressed intensively in the immune system of the clam and the MyD88 gene could recognize the Gram-negative bacterium. |
| Key words: Cyclina sinensis Vibrio anguillarum transcriptome library myeloid differentiation factor 88 real-time PCR |
