上海海洋大学学报2024，Vol.33Issue(6)：1299-1312,14.DOI:10.12024/jsou.20240304466
半滑舌鳎vstm2a基因的响应细菌感染的表达模式和功能
The expression pattern and function of vstm2a gene in response to bacterial infection in Cynoglossus semilaevis
摘要
Abstract
In order to explore the expression characteristics and protein function of vstm2a gene in response to bacterial infection in Cynoglossus semilaevis,in this study,the relative expression characteristics of vstm2a in immune tissues(gill,liver,spleen,intestine and kidney)of Cynoglossus semilaevis after infection with Vibrio harveyi,Vibrio anguillarum,Vibrio alginolyticus,Edwardsiella tarda and Streptococcus agalactiae were detected by real-time fluorescence quantitative PCR(RT-qPCR).The prokaryotic expression recombinant plasmid of vstm2a gene was constructed and the recombinant protein was obtained.The Oxford cup method,bacterial protein binding experiment and siRNA silencing were used to explore the function of vstm2a protein in bacterial invasion.The results showed that the vstm2a protein had a transmembrane domain and an Ig domain,which functions on the cell surface;The expression of vstm2a gene in related immune tissues was significantly increased at 6 h,48 h and 72 h after infection with Vibrio harveyi,Vibrio anguillarum,Vibrio alginolyticus,Edwardsiella tarda and Streptococcus agalactiae.After incubation with bacteria,the vstm2a recombinant protein was co-precipitated with bacteria by low-speed centrifugation.Bacteriostatic experiments showed that it significantly inhibited the growth of bacteria,and after siRNA reducing the vstm2a gene,it inhibited the expression of cytokines IL-6 and IL-10.This study demonstrates that vstm2a protein can respond to bacterial invasion,recognize and bind bacteria,and play an immune defense role against pathogen infection by inhibiting bacterial growth and activating pro-inflammatory cytokines.It is an important immune protein.This study is helpful to analyze the mechanism of bacterial adhesion to cells or fish resistance to bacterial invasion,and provides a theoretical basis for the future selection of resistant varieties by molecular means.关键词
半滑舌鳎/免疫/vstm2a/跨膜蛋白/细菌结合Key words
Cynoglossus semilaevis/immunization/vstm2a/transmembrane protein/bacterial binding分类
农业科技引用本文复制引用
林梦娇,陈亚东,刘洋,王磊,张天时,史萌,陈松林..半滑舌鳎vstm2a基因的响应细菌感染的表达模式和功能[J].上海海洋大学学报,2024,33(6):1299-1312,14.基金项目
山东省重点研发计划(2021LZGC028) （2021LZGC028）
山东省重点研发计划住鲁院士团队支持项目(2023ZLYS02) （2023ZLYS02）
国家重点研发计划(2022YFF1000303) （2022YFF1000303）
国家海水鱼产业技术体系半滑舌鳎岗位项目(CARS-47-G03) （CARS-47-G03）
中国水产科学研究院创新团队项目(2023TD20) （2023TD20）
山东省泰山学者攀登计划(2017) （2017）
