| 摘要: |
| 补体系统作为先天免疫的重要组成部分，是一种复杂的限制性蛋白水解系统，其在免疫系统中发挥着重要的防御作用。为分析马氏珠母贝补体系统的组成及作用机制，使用血细胞样品进行了全长转录组测序建库、基因比对、功能注释，共挖掘到212个潜在补体样组分相关基因。补体样组分基因经同源性比对和结构域检测分析表明，检索到的基因分别编码89个含C1q结构域蛋白、57个C型凝集素蛋白、33个纤维胶凝蛋白、11个纤维蛋白原相关蛋白、8个甘露糖结合型凝集素关联丝氨酸蛋白酶、2个含硫酯蛋白（1个C3分子，1个TEP分子）、1个补体受体、2个补体因子、9个丝氨酸蛋白酶。随机选择12个补体相关基因，使用溶藻弧菌刺激前后的血细胞样品进行实时定量PCR检测其表达水平，结果显示C1q（C1q domain containing protein）、C-lectin、MBL（mannose-binding lectin）、ficolin、MASP（mannan-binding lectin serine protease）等基因均呈现出显著差异表达，表明马氏珠母贝补体系统是一个复杂的多组分效应系统，且可能通过凝集素途径或类似于凝集素途径激活补体系统的免疫作用。研究结果为进一步验证马氏珠母贝中存在的原始补体系统提供了分子生物学证据，同时对深入了解马氏珠母贝免疫防御机制，丰富和发展海洋无脊椎动物免疫学内容也具有重要理论意义。 |
| 关键词: 马氏珠母贝 全长转录组 补体组分 生物信息学分析 实时定量PCR |
| DOI：10.11693/hyhz20210700158 |
| 分类号:Q789 |
| 基金项目:广东省自然科学基金-面上项目，2019A1515011875号；广东省省级科技计划项目（国际科技合作领域），2019A050510044号；2021年度广东省普通高校特色创新类项目，2021KTSCX044号；广东海洋大学2019年“冲一流”省财政专项资金建设项目；国家级大学生创新创业训练计划项目，CXXL2020002号；广东海洋大学“南海学者计划”（2017年度）；广东海洋大学2021年度本科生创新团队项目，CXTD2021001号。 |
附件 |
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| IDENTIFICATION AND ANALYSIS OF COMPLEMENT-LIKE COMPONENTS IN PINCTADA FUCATA BASED ON THE FULL-LENGTH TRANSCRIPTOME SEQUENCE DATASETS |
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WANG Jing, LI Gui-Ying, LIN Kai-Qi, OU Jing, ZHANG Shu-Ping, WANG Zhong-Liang
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Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China
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| Abstract: |
| The complement system in animals is a complex and restricted proteolytic system, and plays a defensive role in the immune system. To explore the composition and mechanism of the complement system in Pinctada fucata, full-length transcriptome sequencing, gene alignment, and functional annotation were performed. Homology comparison and conserved domains analysis showed a total of 212 genes of potential complement components were found in P. fucata, including 89 C1q domain-containing proteins, 57 C-type lectins, 33 ficolins and 11 fibrinogen-related proteins, 8 mannan-binding lectin-associated serine proteases, 2 thioester-containing proteins (1 C3 and 1 TEP), 1 complement receptor, 2 complement factors and 9 serine proteases. Real-time PCR analysis showed that the expression levels of C1q, C-lectin, MBL, ficolin, and MASP were up-regulated significantly after Vibrio alginolyticus challenge, indicating that the complement system of P. fucata is a complex multi-component effective system, and lectin pathway or lectin-like pathway may be used to active the complement system with the immune responses. This study provided molecular evidence for further understanding on the original complement system and the immune defense mechanism of P. fucata, as well as the marine invertebrate immunology. |
| Key words: Pinctada fucata full-length transcriptome sequence complement components bioinformatical analysis real-time PCR |
