马岗鹅TLR4基因的克隆及组织表达分析

Molecular Cloning and Expression Patterns of TLR4 Gene of Magang Goose

为了解马岗鹅天然免疫受体TLR4基因特点及该基因在马岗鹅脏器的表达差异,研究扩增马岗鹅脾脏中的TLR4基因并测序,分析和预测TLR4基因核酸和蛋白特征,应用real-time PCR检测TLR4 mRNA在马岗鹅体内脏器的表达差异。结果显示:马岗鹅的TLR4基因大小为2 611 bp,编码氨基酸843个,分子质量为96.17 ku,理论等电点为7.55,含有30个氨基酸组成的信号肽,蛋白整体表现为疏水性;相似性分析和系统发育树分析表明,马岗鹅的TLR4基因与绿头鸭、原鸡等禽类动物有着较高的相似性;TLR4基因在马岗鹅的脾、法氏囊、胸腺、肺、肠等富含淋巴组织或细胞的器官中相对转录量较高。研究结果为进一步探讨马岗鹅的免疫机制奠定了基础。

To understand the characteristics and distribution of TLR4 gene in Magang goose, TLR4 gene in spleen of Magang goose was amplified and sequenced. The characteristics of TLR4 protein were analyzed and predicted. The real-time PCR was used to detect the distribution of TLR4 gene in Magang goose. The length of TLR4 gene of Magang goose was 2 611 bp, it could encode 843 amino acids with a molecular mass of 96.17 ku. The theoretical isoelectric point was 7.55 and contained a signal peptide of 30 amino acids. Protein prediction indicated that the whole protein of encoded TLR4 showed hydrophobicity. Sequence homology analysis and phylogenetic tree analysis indicated that TLR4gene of Magang goose had higher similarity with poultry such as Anas platyrhynchos and Gallus gallus with high conserved pattern. The result of real-time PCR showed that the relative transcription of TLR4 gene was higher in spleen, bursa, thymus, lung, intestine, which had abundant of lymphoctyes or lymphoid tissue. This study laid the foundation for the further study of the immune mechanism of Magang Goose.