摘要： Toll样受体是参与机体天然免疫的一类模式分子，可以识别多种病原体相关分子模式，快速启动有效的免疫应答，在机体的抗感染免疫中发挥重要的作用。为了深入探讨鸡Toll样受体的免疫功能，为禽类相关疾病的预防和控制提供依据，试验通过RT-PCR技术克隆海兰褐鸡TLR15基因的全长阅读框，同时将其胞外区片段插入载体pET32a并导入大肠杆菌BL21（DE3）中进行优化表达，经SDS-PAGE和Western Blot法检测融合蛋白，并将融合蛋白作为免疫原免疫昆明鼠后制备多克隆抗体，同时采用免疫组织化学的方法，对chTLR15的组织表达进行检测。结果表明，海兰褐鸡TLR15编码基因全长2 607 bp，编码868个氨基酸残基，其中胞外区1 962 bp，重组蛋白分子量约为72 ku；免疫小鼠可以产生特异性的抗体，抗体效价为1∶ 10 000；免疫组化检测结果显示，chTLR15主要在脾脏中表达，在肝脏和肺部表达较少。研究结果可为禽类TLR15相关的免疫调节机制分析提供物质基础，也可为后续抗感染生物制剂的开发奠定基础。

关键词: 海兰褐鸡, TLR15, 表达, 序列分析

Abstract: Toll-like receptor that involve innate immunity is one kind of model molecule, and it can recognize a wide range of pathogen-associated molecular patterns and has important effects on anti-infectious immunity. The TLR15 gene was cloned from the Hy-Line brown chicken by RT-PCR, the extracellular region of TLR15 gene was expressed in prokaryotic cells.The fusion protein was detected by SDS-PAGE and Western Blot assay, and were used to immunize Kunming-mice to prepare polyclonal antibodies. Then the expression of chTLR15 in the main organs was detected by immunohistochemical method. The results showed that the open reading frame of Hy-Line brown chicken TLR15 gene was composed of 2 607 bp in length encoding 868 amino acids, the extracellular region was 1 962 bp, the molecular weight of recombinant protein was regarding 72 ku, and the fusion protein was used for producing the antiserum in mice with the antibody titer 1:10 000. TLR15 protein was mainly expressed in the spleen, but less in liver and lung. The study would provide basic materials for the molecular study on poultry TLR15, and construct a basis for the development of anti-infectious biological agents.

Key words: Hy-Line brown chicken, Toll-like receptor15(TLR15), Expression, Sequence analysis

中图分类号: 

• Q78

• S831