Cathepsin B在感染才女虫虾夷扇贝中表达规律的研究  

Study on the expression pattern of Cathepsin B in Yesso scallops(Patinopecten yessoensis)infected with Polydora

作　　者：杭雲娜 孙红妍 唐安琪 毛俊霞 田莹 王许波 郝振林 丁君 常亚青 HANG Yunna;SUN Hongyan;TANG Anqi;MAO Junxia;TIAN Ying;WANG Xubo;HAO Zhenlin;DING Jun;CHANG Yaqing(Key Laboratory of Mariculture&Stock Enhancement in North China s Sea,Ministry of Agriculture and Rural Affairs,College of Fisheries and Life Sciences,Dalian Ocean University,Dalian 116023,China)

机构地区：[1]大连海洋大学水产与生命学院,农业农村部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《大连海洋大学学报》2025年第1期55-64,共10页Journal of Dalian Ocean University

基　　金：辽宁省科学技术计划面上项目(2023-MS-284);国家重点研发计划“海洋农业与淡水渔业科技创新”重点专项(2022YFD2400302);辽宁省“兴辽英才计划”领军人才项目(XLYC2202001);辽宁省属本科高校基本科研业务费专项资金(LJ212410158060,2024JBYBZ002)。

摘　　要：为探究组织蛋白酶B(Cathepsin B,CatB)在虾夷扇贝(Patinopecten yessoensis)应答才女虫(Polydora)侵染中的分子功能,本研究进行了虾夷扇贝CatB基因(PyCatB)的基因组鉴定、序列特征和基因表达分析,解析了感染才女虫虾夷扇贝中PyCatB的酶活性变化特征和基因表达变化规律。结果表明:PyCatB基因全长为9 267 bp,包含9个外显子,8个内含子;其cDNA长度为1 219 bp, 5′UTR长度为103 bp, 3′UTR长度为99 bp;开放阅读框(ORF)长度为1 017 bp,编码338个氨基酸;PyCatB预测含有N端信号肽、前肽Propeptide C1和肽酶peptidase C1A cathepsin B 3个保守功能域;此外还具有催化三联体结构(Cys116、His284、Asn304)、Gln110氧阴离子孔、GCNGG基序及C194~C205固定残基片段形成的特征性闭塞环,这些结构对PyCatB的催化活性起着重要的作用;系统进化分析表明,各物种CatB的进化地位与生物进化地位一致;酶活性分析显示,感染才女虫虾夷扇贝外套膜组织的CatB酶活性显著高于健康扇贝;基因表达分析表明,除闭壳肌外,PyCatB在健康虾夷扇贝成体各组织中均有不同水平的表达;研究发现,PyCatB基因参与广泛的生理过程;PyCatB在患病虾夷扇贝的外套膜不同区域的表达量均要显著高于健康扇贝,但在健康和患病扇贝血淋巴细胞中的表达无显著性差异,推测PyCatB主要通过外套膜组织参与虾夷扇贝的免疫应答过程进而抵御才女虫的侵染。本研究为虾夷扇贝应答才女虫侵染机制的解析提供了科学参考。Cathepsin B(CatB)is a lysosomal cysteine protease with important immune functions.To investigate the molecular function of CatB in Yesso scallops(Patinopecten yessoensis)infected by Polydora,genomic identification,sequence characterization,and gene expression analyses of the CatB gene were conducted in P.yessoensis(PyCatB),and the changes of PyCatB enzyme activity and gene expression pettern were analyzed.The results showed that the full length of the PyCatB gene was 9267 bp with nine exons and eight introns.The cDNA length of PyCatB was 1219 bp,with a 5′UTR of 103 bp and a 3′UTR of 99 bp,and an open reading frame length of 1017 bp,encoding 338 amino acids.PyCatB was predicted to contain three conserved functional domains;the N-terminal signal peptide;Propeptide C1;and peptidase C1A cathepsin B.Additionally,it also had a catalytic triplet structure(Cys116,His284,Asn304),a Gln110 oxygen anion pore,a GCNGG motif,and a characteristic occluding loop formed by C194/C205 fixed residue fragments,which played important roles in the catalytic activity of PyCatB.Phylogenetic analysis showed that the evolutionary status of CatB was consistent with the species taxonomy.Enzyme activity analysis showed that the enzyme activity of CatB in the mantle tissue of P.yessoensis infected with Polydora was significantly higher than that in healthy scallops.Gene expression analysis showed that PyCatB was expressed at different levels in the various tissues of healthy adult scallops except for the adductor muscle,indicating that PyCatB was involved in a wide range of physiological processes.The expression levels of PyCatB in different mantle regions of infected scallops were all significantly higher than those in healthy scallops,but there was no significant difference between the hemocytes of healthy and infected scallops.It was speculated that PyCatB probably acted mainly through the mantle tissue to participate in the immune response processes of P.yessoensis to resist Polydora infection.This study provides a reference for analyse

关 键 词：虾夷扇贝 才女虫病 组织蛋白酶B 酶活性 表达规律 

分 类 号：S917.4[农业科学—水产科学]