农业生物技术学报2025，Vol.33Issue(7)：1553-1567,15.DOI:10.3969/j.issn.1674-7968.2025.07.012
大口黑鲈PTEN基因的克隆、组织表达及初步功能分析
Cloning,Tissue Expression and Primary Functional Analysis of PTEN Gene in Largemouth Bass(Micropterus salmoides)
摘要
Abstract
Phosphatase and tensin homolog deleted on chromosome ten(PTEN)is the first tumor suppressor gene with bispecific phosphatase activity,which plays an important role in many signaling pathways.To clarify the regulatory role of PTEN in nuclear factor kappa-B(NF-κB),interferon(IFN)and activator protein-1(AP-1)signaling pathways,which are closely related to the immune response in fish,in this study,the ORF region of PTEN gene was cloned from largemouth bass(Micropterus salmoides),the effects of pathogen-associated molecular patterns Nocardia seriolae,Largemouth bass virus(LMBV),Poly I∶C challenge on PTEN gene expression in largemouth bass were detected by qPCR in vivo.To investigate the subcellular localization of PTEN,eukaryotic expression plasmid pCMV-Flag-PTEN was constructed,and the effect of PTEN overexpression on NF-κB,AP-1 and IFN promoter activation was investigated.Protein sequence analysis showed that PTEN encoded a 439-amino acid protein,including a PTPc domain and a PTEN_C2 domain.In the phylogenetic tree,the PTEN protein of largemouth bass was clustered with other fish.The results of qPCR showed that PTEN was widely expressed in many tissues,and highly expressed in spleen and gill.After LMBV and Poly I∶C stimulation,the expression of PTEN decreased first and then increased or increased first and then decreased.The results of subcellular localization showed that PTEN was distributed in the cytoplasm and nucleus of HEK-293T cells.Double luciferase activity assay showed that overexpression of PTEN could enhance AP-1 activation and inhibit the activity of NF-κB and IFN promoter.These results suggested that PTEN played an important regulatory role in the innate immune response against bacteria and viruses by activating AP-1 and inhibiting NF-κB and IFN signaling pathways.This study provides basic data for the clarify of immune response mechanism of largemouth bass against disease,and provides theoretical basis for the development of antibacterial vaccine and the breeding of disease-resistant varieties(lines).关键词
大口黑鲈/第10号染色体上缺失的磷酸酶与张力蛋白同源物基因(PTEN)/基因克隆/组织表达/亚细胞定位/双荧光素酶活性分析Key words
Micropterus salmoides/Phosphatase and tensin homolog deleted on chromosome ten(PTEN)/Gene clone/Tissue expression/Subcellular localization/Dual luciferase activity assay分类
水产学引用本文复制引用
李佳鑫,高风英,董浚键,孙成飞,张赫桐,朱芷琳,王庆奎..大口黑鲈PTEN基因的克隆、组织表达及初步功能分析[J].农业生物技术学报,2025,33(7):1553-1567,15.基金项目
中国水产科学研究院基本科研业务费(2023TD95) （2023TD95）
现代农业产业技术体系专项资金(CARS-46) （CARS-46）
