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引用本文:钟爱华,储张杰,王伟洪,牟毅.泥蚶(Tegillarca granosa)血细胞转录组序列中免疫相关基因的结构与mRNA表达量分析.海洋与湖沼,2020,51(2):354-363.
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泥蚶(Tegillarca granosa)血细胞转录组序列中免疫相关基因的结构与mRNA表达量分析
钟爱华, 储张杰, 王伟洪, 牟毅
浙江海洋大学水产学院 舟山 316022
摘要:
贝类血细胞直接参与异物吞噬和包囊作用,还在伤口修复、炎症反应过程中发挥重要作用,是机体免疫防御的承担者。本研究采用RNA-seq测序技术研究了鳗弧菌感染前后泥蚶血细胞的转录组序列,通过序列组装,获得了α-葡萄糖苷酶(GAA)、α-甘露糖苷酶(LAMAN)、芳基硫酸酯酶B(ASB)、天冬酰胺氨基葡萄糖酶(AGA)、整合素β3亚基(ITB3)、热休克蛋白9(HSPA9)和Toll相互作用蛋白(TOLLIP)序列,利用Orffinder获得了其编码蛋白序列,并对蛋白序列进行了理化性质、二级结构、保守结构域和三级结构分析。结果表明:7种蛋白质二级结构均含有α螺旋、β折叠及无规则卷曲,泥蚶和人类ITB3和TOLLIP具有相同的保守结构域,Swiss-model同源建模获得的三级结构与模板同源性均超过35%(TOLLIP除外)。鳗弧菌刺激后,GAA、LAMAN、ASB、AGA和HSPA9的表达量显著增加,ITB3和TOLLIP表达量变化不明显。该研究补充和完善了对泥蚶免疫相关基因的认识,并为进一步开展泥蚶抗病机理研究提供了重要的理论依据。
关键词:  泥蚶  血细胞  转录组测序  免疫基因
DOI:10.11693/hyhz20191200251
分类号:S917.4
基金项目:浙江省"水产"一流学科开放课题,20160012号;国家自然科学基金项目,31700322号。
ANALYSIS OF THE STRUCTURE AND EXPRESSION OF IMMUNE-RELATED GENES IN BLOODY CLAM TEGILLARCA GRANOSA BASED ON HAEMOCYTE TRANSCRIPTOME SEQUENCING
ZHONG Ai-Hua, CHU Zhang-Jie, WANG Wei-Hong, MOU Yi
School of Fishery, Zhejiang Ocean University, Zhoushan 316022, China
Abstract:
The haemocytes of shellfish are directly involved in the phagocytosis and encapsulation of foreign materials. They also play an important role in the process of wound repair and inflammatory response. They are the undertaker of immune defense. RNA sequencing technology was used to study the transcriptome of haemocytes from bloody clam Tegillarca granosa. By sequence assembly, the sequences of α-glucosidase (GAA), α-mannosidase (LAMAN), arylsulfatase B (ASB), asparagine glucosaminase (AGA), integrin β3 subunit (ITB3), heat shock protein 9 (HSPA9) and toll interaction protein (TOLLIP) were obtained. The encoded proteins were obtained by Orfinder. The physical and chemical properties, secondary structure, conserved domain, and tertiary structure were analyzed. The results showed that the secondary structures of the seven proteins contained α-helix, β-fold, and irregular curl. The ITB3 and TOLLIP conserved domains of clam and human are similar. The homology of the tertiary structure and template from Swiss-Model was more than 35% (except TOLLIP). After stimulation by Vibrio anguillarum, the expression of GAA, LAMAN, ASB, AGA, and HSPA9 increased significantly, while the expression of ITB3 and Tollip did not change significantly. This study complemented and improved the understanding of immune related genes, and provided important information for further research on the disease resistance mechanism of T. granosa.
Key words:  bloody clam Tegillarca granosa  haemocyte  RNA sequencing  immune-related gene
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