China Animal Husbandry and Veterinary Medicine ›› 2023, Vol. 50 ›› Issue (7): 2606-2616.doi: 10.16431/j.cnki.1671-7236.2023.07.002

• Biotechnology • Previous Articles     Next Articles

Cloning and Tissue Expression Specificity Analysis of Interferon Regulatory Factor 7 Gene in Ducks

DENG Zhichao1,2, ZHANG Cheng1,2, LUO Pei1,2, ZHAN Xiaozhi1,2, JIANG Danli1,2, YANG Youtian3, LIU Wenjun1,2   

  1. 1. Zhongkai University of Agriculture and Engineering, Guangzhou 510230, China;
    2. Guangdong Province Key Laboratory of Waterfowl Healthy Breeding, Guangzhou 510230, China;
    3. Foshan University, Foshan 528225, China
  • Received:2022-11-27 Published:2023-06-30

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Abstract: 【Objective】 The purpose of this study was to obtain the eukaryotic expression vector of CDS of interferon regulatory factor 7 (IRF7) gene in Cherry Valley ducks, to study the structure and biological characteristics of IRF7 protein, and to understand the expression level of IRF7 gene in different tissues, so as to lay a foundation for further study of duck innate immune system.【Method】 The CDS of IRF7 gene in Cherry Valley ducks was amplified by RT-PCR, and the eukaryotic expression plasmid was constructed.The protein expression was detected by Western blotting.The effect of Poly(I:C) on the nuclear translocation of IRF7 protein was observed by fluorescence microscope.The nucleotide similarity alignment was performed on the IRF7 gene sequence, and the phylogenetic tree was constructed, and the molecular and structural characteristics of IRF7 protein were analyzed and predicted online by bioinformatics method.Real-time quantitative PCR was used to detect the expression specificity of IRF7 gene in different tissues.【Result】 The CDS of IRF7 gene in Cherry Valley ducks was 1 536 bp and encoded 512 amino acids.Through the eukaryotic expression of the recombinant plasmid, the total protein was extracted and the protein expression was detected by Western blotting.The size of the recombinant protein was about 110 ku.The treatment of duck embryo fibroblasts with Poly (I:C) could cause in the localization of pCMV-C-EGFP-IRF7 in the nucleus.The results of nucleotide similarity showed that the nucleotide sequence similarity of the IRF7 gene between Cherry Valley ducks, Anas platyrhynchos and Gallus gallus was 99.1% and 80.8%, respectively, while the similarity with Xenopus laevis was only 41.8%.The phylogenetic tree results showed that Cherry Valley ducks had the closest genetic relationship with Anas platyrhynchos and Gallus gallus, and the most distant genetic relationship with Xenopus laevis.IRF7 protein was a structurally unstable hydrophobic protein, mainly composed of alpha helix, beta turn and random coil.IRF7 gene was expressed in various tissues, with the highest expression in gallbladder and the lowest in lung.【Conclusion】 The size of the CDS region of IRF7 gene in Cherry Valley ducks was 1 536 bp, encoding 512 amino acids.The IRF7 protein was a structurally unstable hydrophobic protein, and Poly (I:C) stimulation could cause nuclear translocation of IRF7.The expression of this gene varies greatly in different tissues.

Key words: interferon regulatory factor 7(IRF7); protein structure; subcellular localization

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