文章摘要
梁箫,刘钰珠,陈珂,李一峰,杨金龙.厚壳贻贝MyD88-4基因的生物学特性及其对沙氏弧菌的免疫应答[J].水产学报,2019,43(11):2347~2358
厚壳贻贝MyD88-4基因的生物学特性及其对沙氏弧菌的免疫应答
Identification of MyD88-4 in Mytilus coruscus and expression changes in response to Vibrio chagasii challenge
投稿时间:2019-01-06  修订日期:2019-03-18
DOI:10.11964/jfc.20190111611
中文关键词: 厚壳贻贝  髓样分化因子88(MyD88)  基因克隆  组织表达  弧菌感染
英文关键词: Mytilus coruscus  myeloid differentiation factor 88 (MyD88)  gene cloning  tissue expression  Vibrio infection
基金项目:国家重点研发计划政府间国际科技创新合作重点专项(2016YFE0131900);国家自然科学基金(41606147,31101885);上海高校水产高峰学科建设项目
作者单位E-mail
梁箫 上海海洋大学, 国家海洋生物科学国际联合研究中心, 上海 201306
上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306
上海海洋大学, 水产科学国家级实验教学示范中心, 上海 201306 
 
刘钰珠 上海海洋大学, 国家海洋生物科学国际联合研究中心, 上海 201306
上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306 
 
陈珂 上海海洋大学, 国家海洋生物科学国际联合研究中心, 上海 201306
上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306 
 
李一峰 上海海洋大学, 国家海洋生物科学国际联合研究中心, 上海 201306
上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306
上海海洋大学, 水产科学国家级实验教学示范中心, 上海 201306 
 
杨金龙 上海海洋大学, 国家海洋生物科学国际联合研究中心, 上海 201306
上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306
上海海洋大学, 水产科学国家级实验教学示范中心, 上海 201306 
jlyang@shou.edu.cn 
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中文摘要:
      为理解髓样分化因子88(myeloid differentiation factor 88, MyD88)基因的生物学特性以及对细菌胁迫的响应,本研究克隆了厚壳贻贝MyD88基因(命名为McMyD88-4)cDNA全长序列,其全长3 930 bp,开放阅读框2 607 bp,编码868个氨基酸。其中,13~109位的氨基酸序列为死亡结构域(death domain,DD),347~481位的氨基酸序列为TIR(toll/interleukin-1 receptor)结构域,TIR结构域包含3个高度保守的区域Box 1、Box 2和Box3;McMyD88-4蛋白的空间结构包含6个α螺旋(α-helix)和4个β折叠(β-sheet)。同源性分析显示,McMyD88-4蛋白序列与长牡蛎MyD88最相似,其一致性和相似性分别为60 %和77 %;其次,与虾夷扇贝、海湾扇贝和菲律宾蛤仔相似度较高,其一致性和相似性分别为40%~51%和58%~67%。系统进化树结果显示,McMyD88-4先与长牡蛎和扇贝聚为一支,然后与黑腹果蝇聚为一支,脊椎动物单独聚为一支。实时荧光定量PCR(qRT-PCR)检测发现,McMyD88-4基因在厚壳贻贝各组织和器官中均有表达,其中在外套膜和鳃中的表达量最高,而血细胞中表达量最低。厚壳贻贝经沙氏弧菌感染后,McMyD88-4基因表达量在免疫相关组织中急剧上升,分别在感染后3和6 h达到峰值,且在消化腺中的上调水平显著高于鳃和外套膜。研究表明,McMyD88-4在厚壳贻贝抵御外界病原体侵染过程中,尤其是弧菌感染方面发挥重要作用。
英文摘要:
      Myeloid differentiation factor 88 (MyD88) is a crucial adaptor protein in the Toll-like receptor (TLR) signaling pathway and plays an important role in signal transmission. In this study, the full-length cDNA sequence of a MyD88 gene from Mytilus coruscus (McMyD88-4) was cloned. Its cDNA sequence is 3 930 bp, with a 2 607 bp open reading frame encoding a protein of 868 amino acids. Amino acids at position of 13-109 are the dead domain, and amino acids at position of 347-481 are the Toll/interleukin-1 receptor (TIR) domain, which contains three highly conserved regions Box 1, Box 2 and Box3. The three-dimensional structure of McMyD88-4 contains six α-helix and four β-sheets. According to the comparison with known MyD88 amino acids sequence, the putative protein of McMyD88-4 was most similar to that of Crassostrea gigas, with the identity and similarity of 60% and 77% respectively. Then it was more similar to Patinopecten yessoensis, Argopecten irradians, Ruditapes philippinarum, with 40%-51% of identity and 58%-67% of similarity. The result of the phylogenetic tree showed that McMyD88-4 of M. coruscus firstly clustered with C. gigas and scallops, and then with Drosophila melanogaster, while vertebrates formed a separate cluster. qRT-PCR revealed that McMyD88-4 was expressed in all tissues and organs examined. The highest expression was found in the mantle and gill, while the lowest expression was found in haemocytes. After infection by Vibrio chagasii, the expression of McMyD88-4 was up-regulated sharply in immune-related tissues, and reached the peak at 3 h and 6 h, and the up-regulation level in digestive gland was significantly higher than those in gill and mantle. The present results showed that McMyD88-4 plays an important role during the immune response of M. coruscus against external pathogens, especially Vibrio infections.
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