| 摘要: |
| 鲍以富含硫酸化多糖（如岩藻多糖、琼脂糖等）的大型藻类为食，其消化过程依赖硫酸酯酶对硫酸酯键的水解作用，但目前关于硫酸酯酶在动物消化系统中的功能演化机制尚未明晰。本研究分析了皱纹盘鲍（Haliotis discus hannai）基因组，发现其染色体上存在一个由4个芳基硫酸酯酶B（arylsulfatase B, ARSB）基因家族成员组成的串联重复基因簇。同源比对和系统发育分析进一步揭示该基因簇在鲍属其他3个物种中均同样存在，表明这种串联复制事件很可能发生于鲍共同祖先的基因组中，并在后代物种中得以保留。对其序列分析显示，该基因簇成员在基因结构上存在差异，且保守位点区域发生氨基酸突变。荧光定量PCR结果表明，复制基因在不同藻类喂养的皱纹盘鲍中存在表达差异。为进一步解析硫酸酯酶复制基因的功能特征，本研究克隆了HdARSB2基因与鲍甲酰甘氨酸生成酶基因HdFGE，并导入原核细胞进行共表达，成功获得了可溶性重组蛋白，但未在上清液中检测到硫酸酯酶活性。原位杂交分析表明HdARSB2基因特异性表达于鲍肝胰腺中某个功能集中的腺体。本研究通过解析硫酸酯酶基因的演化轨迹及表达模式，为理解鲍类物种适应复杂食源环境和多变海洋生态系统的分子机制提供了新线索，同时为皱纹盘鲍养殖、育种工作和硫酸酯酶的开发利用提供一定的理论支持。 |
| 关键词: 皱纹盘鲍 芳基硫酸酯酶 基因复制 功能分化 |
| DOI： |
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| 基金项目:山东省重点研发项目(2024LZGCQY003),泰山学者青年专家项目 (tsqn202211250)。 |
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| The Replication and Functional Differentiation of an Arylsulfatase B Gene Cluster in Haliotis discus hannai |
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Yao Yibing1, Xie Yusu2, Liu Xiao1, Xu Fei3
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1.Zhejiang Ocean University;2.Institute of Oceanololgy, Chinese Academy of Sciences;3.Institute of Oceanology, Chinese Academy of Sciences
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| Abstract: |
| Abalone feed on macroalgae rich in sulfated polysaccharides (such as fucoidan and agarose). Their digestive process depends on the hydrolysis of sulfate bond by sulfatase, but the functional evolution mechanism of sulfatase in animal digestive system is not clear at present. This study analyzed the genome of the Pacific abalone (Haliotis discus hannai) and identified a tandemly repeated gene cluster composed of four arylsulfatase B (ARSB) gene family members on its chromosome. Homologous alignment and phylogenetic analysis further revealed that the gene cluster existed in three related abalone species, indicating that this tandem replication event probably occurred in the genome of the common ancestor of abalone and was preserved in the offspring species. The analysis of the gene cluster structure shows that there are some differences in gene structure and amino acid mutations in conserved sites. The results of qPCR also show that the replicated genes are expressed differently in Haliotis discus hannai when fed with different macroalgae. To further investigate the functional characteristics of the duplicated sulfatase genes, this study cloned HdARSB2 and formylglycine-generating enzyme gene HdFGE from abalone, co-expressed them in a prokaryotic system, and successfully obtained soluble recombinant proteins. However, no sulfatase activity was detected in the supernatant. In situ hybridization analysis showed that HdARSB2 gene specifically expressed in a gland with concentrated function in the hepatopancreas of abalone. By analyzing the evolution and expression of sulfatase genes, this study reveals new clues about the molecular mechanisms enabling abalone to adapt to diverse food sources and changing marine environments, while also offering theoretical support for abalone aquaculture, selective breeding, and the potential utilization of sulfatases. |
| Key words: Haliotis discus hannai arylsulfatase gene duplication functional divergence |