卢氏鸡主要白细胞介素的克隆分析与il21的原核表达-cloning and analysis of main interleukin in lushi chicken and prokaryotic expression of il21.docxVIP

摘要 摘要 摘要 摘要 PAGE PAGE IV III III 鸡 IL-21 蛋白不仅能明显刺激鸡外周血淋巴细胞的增殖，而且还能促进外周血淋 巴细胞分泌 IL-2 和 IFN-γ。 本课题了解了卢氏鸡与其它品种禽类和哺乳动物白细胞介素在分子结构上的 异同，而且首次构建了鸡 IL-21 原核表达系统，实现了鸡 IL-21 重组成熟蛋白的 高效表达并初步测定了其生物学活性，为今后研究和利用家禽细胞因子和进一步 研究鸡 IL-21 的结构和生物学功能奠定了良好的基础。 关 键 词：鸡；白细胞介素；IL-21；原核表达 论文类型：基础研究 Subject: Cloning and Bioinformatic Analysis of Main Interleukins from Lushi Chicken and Prokaryotic Expression of IL-21 Specialty: Basic Veterinary Medicine Name: Xu Zhang Supervisor: Professor Chunjie Zhang ABSTRACT Interleukin (IL) plays an important role in the immune responses, inflammatory and hematopoietic systems, and it is also of great value in controling disease. In recent years, the research of poultry interleukin developped rapidly, and it involved many breeds of birds, but few interleukins of each species were studied, and the research was not systematic and comprehensive. Lushi chicken, known as its unique green eggs, is one of excellent chicken breeds in Henan Province. Currently interleukins of Lushi chicken is not reported in China and other countries. Cloning of Lushi chicken interleukin genes and studying their biological activity systematically was very important not only for chicken immune system and immune mechanism, but also for immunology and biological evolution comparison with human and another mammalian. IL-21 has a wide range of biological functions. It can stimulate T cells, B cells, NK cell to proliferation, and enhance the cytotoxic activity of NK cells. It has an important role in host defense and anti-tumor function. Therefore, IL-21 has great potential applications for it as a new immunomodulatory agent and adjuvant. In this study, IL-1β, IL-2, IL-3, IL-4, IL-9, IL-12B, IL-15, IL-17, IL-18, IL-21 were cloned from total RNA of Lushi chicken spleen lymphocytes stimulated by ConA via RT- PCR techique. Then by using a variety of softwares and online servers, the signal peptide, protein molecul