鸡RACK1基因克隆与生物信息学分析及其在鸡不同组织中的表达分析OA

This study aims to investigate the biological characteristics of the chicken receptor for activated C kinase 1(RACK1)protein,assess its role in infectious bursal disease virus(IBDV)replication,and examine its expression pat-terns across various chicken tissues.The RACK1 gene from chickens was cloned,and structural and functional predic-tions of the encoded protein were performed using various bioinformatics tools.A eukaryotic expression vector carrying chicken RACK1 gene was constructed,and its expression was verified by Western blot.The effect of RACK1 overexpres-sion on IBDV replication was also evaluated.Furthermore,the relative expression levels of RACK1 in different chicken tissues were determined by qRT-PCR.The study revealed that chicken RACK1 consisted of 317 amino acid residues,with a molecular mass of approximately 34.87 ku.The protein lacks transmembrane domains and signal peptide but contained seven typical WD40 domains.Its secondary structure was primarily composed of random coils(36.59％)and extended strands(45.11％).Protein-protein interaction predictions suggested that RACK1 may interact with ribosomal proteins such as RPL38,RPL27A,RPL23,RPLP0,and RPS2.Phylogenetic analysis demonstrated that RACK1 was highly conserved across several species.The pCAGGS-HA-chRACK1 eukaryotic expression vector was successfully constructed and con-firmed to be expressed in cells.In an IBDV infection model,RACK1 overexpression significantly enhanced viral replica-tion.qRT-PCR analysis of different chicken tissues revealed that RACK1 expression was highest in the heart and lowest in the spleen.In conclusion,this study characterizes the molecular structure and tissue distribution of chicken RACK 1 and demonstrates its positive regulatory role in IBDV replication,providing a theoretical basis for further research into RACK1 function in avian viral infections and immune responses.