摘要跨膜蛋白9B(transmembrane protein 9 domain family member B, TMEM9B)是近年来被发现的一种多功能糖蛋白，研究表明TMEM9B蛋白在多条信号通路中发挥关键调控作用。而目前，关于该基因的表达和功能特性研究报道较少。本研究选择海兰褐鸡(Gallus gallus)、爱拔益加肉鸡(arbor acres plus broiler, AA+肉鸡)和三黄鸡为研究对象，通过克隆不同品系鸡Tmem9B基因的全长编码序列并对比分析，同时利用实时荧光定量PCR(quantitative real time polymerase chain reaction, qRT-PCR)技术定量鉴定Tmem9B基因在3个品系(AA+肉鸡、三黄鸡和海兰褐鸡)正常雏鸡和大肠杆菌(Escherichia coli)脂多糖(lipopolysaccharide, LPS)诱导的炎症模型雏鸡(AA+肉鸡和海兰褐鸡)的表达变化，研究Tmem9B基因在不同品系雏鸡间的表达活性与功能特性。实验结果表明，三黄鸡Tmem9B基因编码序列存在1个SNP位点，该基因在不同品系鸡间高度保守。组织表达谱分析显示Tmem9B基因在AA+肉鸡、三黄鸡和海兰褐雏鸡的多种组织中均有表达，但是不同品系雏鸡间略有差异。有趣的是，Tmem9B基因在3个品系雏鸡的脑组织中均具有高转录活性。在LPS诱导的炎症模型雏鸡中，Tmem9B基因在免疫相关组织中的转录活性均显著上调。本研究表明，鸡Tmem9B可能是在鸡的生长发育和炎症反应等生物学过程中具有重要作用的一个多功能基因。本研究结果为进一步探索Tmem9B基因的功能与特性提供了理论依据。
Abstract：TMEM9B (transmembrane protein 9 domain family, member B), found in recent years, is a multifunctional transmembrane glycoprotein. Several studies have shown that TMEM9B protein plays key roles in multiple signaling pathways. However, the studies on the expression and functional properties of Tmem9B gene are seldom reported. In this study, Hy-Line brown chicken (Gallus gallus), Arbor acres plus broiler (AA+ broiler) and Sanhuang broiler were selected. The full-length coding sequences of Tmem9B gene were cloned from different chicken breeds and analyzed using bioinformatics tools. The expressional changes of Tmem9B gene in three breeds of normal chicken (AA+ broiler, Sanhuang broiler and Hy-Line brown chicken) and Escherichia coli lipopolysaccharide(LPS)-induced chicken inflammation model (AA+ broiler and Hy-Line brown chicken) were determined respectively to study the expressional activities and functional characteristics using quantitative real time polymerase chain reaction (qRT-PCR) technique. The results showed that there was only one SNP (single nucleotide polymorphism) site in the coding sequence of Sanhuang broiler Tmem9B gene, which meant that the gene was highly conserved among different chicken breeds. Tissue expression profile analysis showed that Tmem9B gene was all expressed in various tissues of AA+ broiler, Sanhuang broiler and Hy-Line brown chicken, but there were a little differences between different chicken breeds. Interestingly, the Tmem9B gene had high transcriptional activity in brain of all the three breeds. The expressions of Tmem9B gene of immune-related tissues were significantly up-regulated in the inflammation model chicken induced by LPS (lipopolysaccharide). This study showed hat Tmem9B may be a multifunctional gene that played important roles in many chicken biological processes, such as growth, development and inflammatory response. This study may provide theoretical basis for further explanation of the function and characteristics of Tmem9B gene.
