文献类型：期刊文献

中文题名：利用酵母双杂交系统筛选与鉴定石斑鱼EcBAG3互作因子

英文题名：Screening and Identifing the Interacting Proteins of Grouper(Epinephelus coioides)EcBAG3 Using Yeast Two-hybrid System

作者：蔡佳[1,2];梁振宇[1];黄瑜[1];鲁义善[1];施钢[1];简纪常[1]

机构：[1]广东海洋大学水产学院,广东省水产经济动物病原生物学及流行病学重点实验室,广东省水产经济动物病害控制重点实验室,南方海洋科学与工程广东省实验室(湛江),湛江524088;[2]广西科学院,广西北部湾海洋研究中心,广西海洋天然产物与组合生物合成化学重点实验室,南宁530007

年份：2022

卷号：38

期号：8

起止页码：77

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD、、CSCD2021_2022、北大核心、CSCD、北大核心2020

基金：国家自然科学基金项目(U20A2065,32002426,32073006);广西自然科学基金(2020GXNSFAA297243);南方海洋科学与工程广东省实验室(湛江)(湛江湾实验室)自主立项项目(ZJW-2019-06);广东海洋大学“冲一流”专项资金项目(231419013,231419017);广东海洋大学“冲一流”与“创新强校工程”科研项目(2019KTSCX056);广西红树林滨海湿地生态保护与可持续利用人才小高地人才资助项目(BGMRC202101)。

语种：中文

中文关键词：石斑鱼;酵母双杂交;BAG3;互作蛋白

外文关键词：grouper(Epinephelus coioides);yeast two hybrid;BAG3;interacting proteins

中文摘要：以参与石斑鱼抗病毒免疫反应的EcBAG3蛋白为研究对象,采用酵母双杂交等技术筛选与鉴定EcBAG3的互作蛋白。首先构建了石斑鱼脾脏细胞的酵母双杂交cDNA文库,文库的容量为5.6×10^(6) CFU,插入片段的平均长度在750 bp以上,重组率为100%。进一步利用此文库筛选EcBAG3的互作因子,初步得到了83个阳性克隆。对这些克隆进行测序与一对一互作验证,最终获得7个候选互作蛋白,功能预测结果显示,互作蛋白涉及免疫调控、转录调节、胁迫响应、信号转导等方面的功能,为进一步解析石斑鱼BAG3在病毒感染中的作用机制奠定了基础。

外文摘要：The EcBAG3 protein involved in the antiviral immune response of grouper(Epinephelus coioides)was used as the research object,and the interacting proteins of EcBAG3 were screened and identified by yeast two-hybrid technology.Firstly,a yeast two-hybrid cDNA library of grouper(Epinephelus coioides)spleen cells was constructed.The capacity of the library was 5.6×106 CFU,the average length of the inserted cDNA fragments was over 750 bp,and the recombination rate was 100%.Furthermore,the interaction factors of Ec-BAG3 were screened from this library and 83 positive clones were initially obtained.Finally 7 candidate interacting proteins were confirmed through sequencing and one-to-one interaction verification.The functional prediction showed that the interacting proteins were involved in immune regulation,transcriptional regulation,stress response,signal transduction,etc.,which laid a foundation to further understand the mechanism of EcBAG3 during viral infection.