Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (8): 77-83.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1368

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

CAI Jia1,2(), LIANG Zhen-yu1, HUANG Yu1, LU Yi-shan1, SHI gang1, JIAN Ji-chang1   

  1. 1. Fisheries College,Guangdong Ocean University,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institutes,Guangdong Southern Marine Science and Engineering Laboratory(Zhanjiang),Zhanjiang 524088
    2. Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry,Guangxi Beibu Gulf Marine Research Center,Guangxi Academy of Sciences,Nanning 530007
  • Received:2021-10-29 Online:2022-08-26 Published:2022-09-14
  • Contact: CAI Jia E-mail:matrix924@foxmail.com

Abstract:

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.

Key words: grouper(Epinephelus coioides), yeast two hybrid, BAG3, interacting proteins