南方农业学报2025，Vol.56Issue(7)：2080-2088,9.DOI:10.3969/j.issn.2095-1191.2025.07.004
叉角厉蝽组织蛋白酶B基因克隆及表达分析
Gene cloning and expression analysis of the cathepsin B in Eocanthecona furcellata
摘要
Abstract
[Objective]The cathepsin B gene of Eocanthecona furcellata was(EfCAB)cloned and its expression was analyzed to investigate the role of EfCAB protein in the life activities of Eocanthecona furcellata,thereby providing a foundation for studying its physiological functions.[Method]The open reading frame(ORF)sequence of EfCAB gene was amplified and cloned by PCR.Bioinformatics software were used to analyze the physicochemical properties,trans-membrane structure,signal peptide,and subcellular localization of EfCAB protein.Comparative alignment and similarity analysis of cathepsin B amino acid sequences among different species were conducted and a phylogenetic tree was con-structed using MEGA 11.0.Real-time quantitative PCR(qPCR)was used to detect the expression characteristics of the Ef-CAB gene in different tissues(malpighian tube,salivary gland,ovary,head,gut,and fat body of female adults)and dif-ferent developmental stages(eggs,1st to 5th instars,and male and female adults).[Result]The ORF region of EfCAB with the theoretical isoelectric point of 6.95.The EfCAB protein was not classified as a transmembrane protein as it did not have a transmembrane structure domain;the first 18 amino acids of the EfCAB protein served as a signal peptide,indicating that it is a secreted protein.The EfCAB protein was primarily localized in the cytoplasm and mitochondria.The highest similarity of amino acid sequence was observed between the EfCAB protein and the cathepsin B of the Haly-omorpha halys(87.0%);phylogenetic analysis results indicated that the Eocanthecona furcellata was most closely related to the Halyomorpha halys.The qPCR results showed that the relative expression of EfCAB gene in the malpighian tube was significantly higher than that in other tissues(P<0.05,the same below),with a relatively high expression in gut but almost no expression in head;the relative expression of EfCAB gene was the highest in eggs,which was significantly higher than that in other developmental stages,with relatively high expression observed in the 3rd instar larvae and female adult stage,and the lowest relative expression in 5th instar larvae stage.[Conclusion]The expression of the EfCAB gene exhibits tissue specificity and stage specificity,suggesting that for Eocanthecona furcellata,the EfCAB gene helps digest prey tissues in the predation process and promote its embryonic development.关键词
叉角厉蝽/组织蛋白酶B/基因克隆/表达特征Key words
Eocanthecona furcellata/cathepsin B/gene cloning/expression characteristics分类
农业科技引用本文复制引用
陈岗,岳茂婷,丁以纾,何文德,吴子云,陈晓,吴国星,高熹..叉角厉蝽组织蛋白酶B基因克隆及表达分析[J].南方农业学报,2025,56(7):2080-2088,9.基金项目
云南省农业基础研究联合专项(202301BD070001-137,202101BD070001-050) （202301BD070001-137,202101BD070001-050）
云南省中青年学术和技术带头人后备人才项目(202205AC160077) （202205AC160077）
云南省烟草公司科技计划项目(2023530000241012) Agricultural Joint Special Project of Yunnan Province(202301BD070001-137,202101BD070001-050) （2023530000241012）
Reserve Talent Project for Young-and Middle-aged Academic and Technical Leaders of Yunnan Province(202205AC160077) （202205AC160077）
Scien-ce and Technology Project of Yunnan Tobacco Company(2023530000241012) （2023530000241012）
