摘要
【目的】深入研究克氏原螯虾(Procambarus clarkia)traf6 like基因参与先天免疫调节的功能。【方法】以Pc-traf6 like基因为研究对象,利用RNAi技术干扰目的基因表达量之后,进行细菌模拟刺激,并且通过实时荧光定量PCR(qRT-PCR)检测细胞凋亡相关基因表达量的变化情况。【结果】干扰Pc-traf6 like基因,通过干扰效果检测,显示48 h时干扰效果良好,可用于后续试验。在目的基因干扰后,进行金黄色葡萄球菌(Staphylococcus aureus)刺激,检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达;在目的基因干扰后,进行鲶爱德华氏菌(Edwardsiella ictaluri)刺激,检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达。【结论】Pc-traf6 like基因被干扰,机体在受到金黄色葡萄球菌及鲶爱德华氏菌刺激后,Bax、Bcl 2和Caspase 3基因的相对表达量上调,推测Pc-traf6 like基因通过调控细胞凋亡参与了先天免疫反应。
【Objective】Role of traf6 like gene in innate immunity regulation of Procambarus clarkia(Pc-traf6 like)was investigated.【Methods】RNAi technology was employed to interfere the expression of Pc-traf6 like gene.Bacterial simulation was performed to determine the changes of apoptosis-related gene expression by real-time quantitative PCR(qRTPCR).【Result】The interference on the gene was effective after 48 h for the subsequent experiments.The interferenceinduced pro-apoptotic gene Bax,anti-apoptotic gene Bcl 2,and apoptotic protease Caspase 3 were upregulated in varying degrees with the stimulation of Staphylococcus aureus or Edwardsiella ictaluri.【Conclusion】After the Pc-traf6 like gene was effectively interfered,the relative expressions of Bax,Bcl 2,and Caspase 3 were upregulated after the P.clarkia was stimulated by either S.aureus or E.ictaluri indicating an involvement of Pc-traf6 like gene in the innate immune response by the regulating apoptosis.
作者
蔺思函
杨兵兵
李倩倩
沈秀丽
杜志强
LIN Sihan;YANG Bingbing;LI Qianqian;SHEN Xiuli;DU Zhiqiang(College of Life Science and Technology,Inner Mongolia University of Science and Technology,Baotou,Inner Mongolia014010,China;Library,Inner Mongolia University of Science and Technology,Baotou,Inner Mongolia 014010,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2022年第10期1245-1249,共5页
Fujian Journal of Agricultural Sciences
基金
内蒙古自治区自然科学基金项目(2020JQ03)
国家自然科学基金项目(32060834)
内蒙古自治区研究生科研创新项目(S20210160Z)
作者简介
蔺思函(1998-),女,硕士研究生,研究方向:无脊椎动物的先天免疫(E-mail:2836019366@qq.com);通信作者:杜志强(1980-),男,博士,教授,研究方向:无脊椎动物的先天免疫(E-mail:nmdzq@163.com)
