关键词: 白来航鸡; STING基因; 克隆; 生物信息学; 表达

Abstract: 【Objective】 In this study,the CDS region sequence of stimulator of interferon genes (STING) gene in White Leghorn chickens was cloned and analyzed by bioinformatics and tissue expression,which laid a foundation for elucidating the role of STING gene in antiviral immune response.【Method】 The CDS region of STING gene in White Leghorn chickens was amplified by PCR and cloned.After sequencing,the similarity of the encoded amino acid sequence of STING gene was compared and the phylogenetic tree was constructed.The physicochemical properties and structural function of STING protein were predicted by bioinformatics,and the expression of STING gene in 14 tissues such as heart and liver of White Leghorn chickens were detected by Real-time quantitative PCR.【Result】 The sequence of the CDS region of STING gene in White Leghorn chickens was 1 140 bp in total length,encoding 379 amino acids.Similarity alignment and phylogenetic tree analysis showed that STING gene in White Leghorn chickens had the highest similarity with Gallus gallus and the closest genetic relationship,and the farthest genetic relationship with Corvus cornix cornix.STING protein in White Leghorn chickens was an acidic,hydrophilic protein with a molecular weight of 42.625 ku,an isoelectric point (pI) of 6.67,an instability coefficient of 69.26, and a fat coefficient of 105.01.STING protein was synthesized mostly on mitochondria and endoplasmic reticulum,contained a transmembrane structure,and no signal peptide.The secondary structure of STING protein included alpha helix (54.62%),extended chain (10.29%),beta turn (3.43%) and random coil (31.66%).Protein interaction analysis showed that there were interactions between STING and NFKB1,DDX41,cGAS,TBK1 and other proteins.The results of Real-time quantitative PCR showed that STING gene was widely expressed in the tissues of White Leghoron chickens,with the highest expression in lung,which was significantly higher than that in other tissues (P＜0.05),and the lowest expression in pectoralis muscle.【Conclusion】 In this study,STING gene in White Leghorn chickens was successfully cloned,the total length of the CDS region was 1 140 bp,encoding 379 amino acids.STING protein in White Leghorn chickens was an acidic,hydrophilic protein with a transmembrane structure.There was the highest expression of STING gene in lung of White Leghoron chickens.The results provided materials for the in-depth study of the function of the protein encoded by STING gene in White Leghorn chickens.

Key words: White Leghorn chickens; STING gene; cloning; bioinformatics; expression