鸡热休克蛋白8基因克隆、生物信息学分析及其在胸腺中表达模式研究

doi:10.16431/j.cnki.1671-7236.2025.01.004

Abstract

Abstract: 【Objective】 The aim of this study was to investigate the biological characteristics of heat shock protein family A member 8 (HSPA8) in chickens and its expression patterns in thymus under stress. 【Method】 In this study,the thymus of Gushi chickens was used as the material,and the CDS region of HSPA8 gene was amplified using PCR and cloned,and its biological and structural characteristics were analyzed by bioinformatics method.The relative expression of HSPA8 gene in different tissues were detected. The models of beak trimming stress and heat stress were constructed,and Real-time quantitative PCR was used to detect the expression of inflammatory factor and HSPA8 gene in thymus of chickens with different stress models. 【Result】 The length of CDS region of chicken HSPA8 gene was 1 941 bp,encoding 646 amino acids,which was 98.92% similar to the gene sequence of Gallus gallus.Phylogenetic tree results of HSPA8 gene sequence showed that Gushi chickens was the closest related to Meleagris gallopavo,followed by mammals,and most distantly related to lower fish.HSPA8 protein was acidic protein with strong stability and hydrophilicity.It didn’t belong to secreted protein and had no transmembrane domain,with multiple phosphorylation and glycosylation modification sites.It mainly functions in the nucleus and had two conserved domains.It interacted with DNAJC5,HSPB1,DNAJA1 and GAK proteins,et al.HSAP8 gene was widely expressed in various tissues and organs of chicks,with the highest expression in pancreas and the lowest expression in pectoral muscle.The results of stress model evaluation showed that beak trimming stress and heat stress resulted in the significant increase of the contents of serum corticosterone,interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (P＜0.05),and decrease of the contents of T cell subsets (CD3⁺) and immunoglobulin G (IgG).The expressions of IL-1β,IL-6,TNF-α and IL-8 genes in thymus were significantly increased on day 1 (P＜0.05) and significantly decreased on day 5 (P＜0.05),while the expressions of IL-1β,IL-6,TNF-α and IL-8 genes were significantly increased in thymus under heat stress (P＜0.05).The expression of HSPA8 gene in thymus were increased significantly on day 1 and day 3 after beak trimming (P＜0.05),decreased significantly on day 5 (P＜0.05),and decreased significantly in the continuous heat stress group (P＜0.05). 【Conclusion】 HSPA8 gene was involved in the stress immune response of chicken thymus,which could be used as a reliable indicator of whether poultry was in the state of stress immunosuppression.The above results provided a reference for further studying the immune regulatory function of HSPA8 gene.

Key words: chickens; HSPA8 gene; bioinformatics; thymus; regulation of stress

CLC Number:

S831

TIAN Huihui, WANG Bingxin, ZHU Zhaoyan, YU Yange, DING Mengxia, TIAN Yadong, JIANG Ruirui, SUN Guirong, HAN Ruili, KANG Xiangtao, YAN Fengbin, GUO Yujie. Cloning,Bioinformatics Analysis of HSPA8 Gene and Its Expression Pattern in Thymus in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 39-51.

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Cloning,Bioinformatics Analysis of HSPA8 Gene and Its Expression Pattern in Thymus in Chickens

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