Department of Animal Science, Guangdong Ocean University, Zhanjiang, 524088, China.
Department of Veterinary Medicine, Guangdong Ocean University, Zhanjiang, 524088, China.
Vet Immunol Immunopathol. 2021 Jun;236:110236. doi: 10.1016/j.vetimm.2021.110236. Epub 2021 Apr 1.
Disease outbreaks heavily impact the economic viability of animal industries. Little is known about the mechanisms of immune system-related diseases in geese. Toll-like receptors (TLRs) play a major role in the anti-inflammatory immunity process in most animal species, but they have not been studied in the Magang goose. To elucidate the role of TLRs, reverse transcription polymerase chain reaction (RT-PCR) and PCR amplification of cDNA ends (Smart RACE) were used to clone the Magang goose TLR5 gene (mgTLR5). The full-length cDNA of mgTLR5 was 2967 bp in length, including a 5'-terminal untranslated region (UTR) of 215 bp, a 3'-terminal UTR of 384 bp, and an open reading frame of 2583 bp that encodes a protein of 860 amino acids. Structurally, mgTLR5 has a toll/interleukin-receptor (TIR) domain, a transmembrane domain, and seven leucine-rich repeats (LRRs) domains. Homology alignment of TLR5 and its TIR domains with other species revealed that mgTLR5 shared 98 % and 81.3 % of sequence similarity with white goose TLR5 and chicken TLR5, respectively. Quantitative RT-PCR showed that the mgTLR5 gene of the goose is widely expressed in all tested tissues, with the highest expression in the kidney and spleen. The increase in NF-κB promoter activity stimulated by flagellin was dependent on mgTLR5 expression in 293 T cells. Salmonella pullorum and flagellin significantly upregulated the expression of TLR5, IL-8, and IL-1 mRNA in peripheral blood mononucleotide cells of Magang goose cultured in vitro. Stimulation by S. pullorum for 24 h upregulated mgTLR5 expression in the cecum and kidney. We conclude that Magang goose TLR5 is a functional TLR5 homologue of the protein in other species and plays an important role in bacterial recognition.
疾病爆发严重影响动物产业的经济活力。人们对鹅的免疫系统相关疾病的机制知之甚少。Toll 样受体 (TLR) 在大多数动物物种的抗炎免疫过程中发挥着重要作用，但在 Magang 鹅中尚未进行研究。为了阐明 TLR 的作用，使用逆转录聚合酶链反应 (RT-PCR) 和 cDNA 末端的 PCR 扩增 (Smart RACE) 克隆了 Magang 鹅 TLR5 基因 (mgTLR5)。mgTLR5 的全长 cDNA 长 2967bp，包括 215bp 的 5'端非翻译区 (UTR)、384bp 的 3'UTR 和 2583bp 的开放阅读框，编码 860 个氨基酸的蛋白质。结构上，mgTLR5 具有 toll/interleukin-receptor (TIR) 结构域、跨膜结构域和七个亮氨酸丰富重复 (LRR) 结构域。TLR5 及其 TIR 结构域与其他物种的同源性比对表明，mgTLR5 与白鹅 TLR5 和鸡 TLR5 的序列相似性分别为 98%和 81.3%。定量 RT-PCR 显示鹅的 mgTLR5 基因在所有检测组织中广泛表达，在肾脏和脾脏中表达最高。鞭毛蛋白刺激 NF-κB 启动子活性的增加依赖于 293T 细胞中 mgTLR5 的表达。体外培养的 Magang 鹅外周血单核细胞中，鸡白痢沙门氏菌和鞭毛蛋白显著上调 TLR5、IL-8 和 IL-1 mRNA 的表达。S. pullorum 刺激 24 小时后，盲肠和肾脏中 mgTLR5 的表达上调。我们得出结论，Magang 鹅 TLR5 是其他物种蛋白质的功能性 TLR5 同源物，在细菌识别中发挥重要作用。
