Abstract: Abstract: Objective To preliminarily analyze the immunogenicity of E． coli expressed chicken Interleukin-10 ( chIL-10) antiserum in stably chIL-10-expressing cell line transfected by chIL-10 gene with signal peptide ( CHOchIL- 10F) and that without signal peptide ( CHO-chIL-10M) ． Method Whole chIL-10 gene was amplified by PCＲ from chicken peripheral blood lymphocytes and cloned into pET-30a ( + ) vector to obtain recombinant plasmid of pET-chIL-10，followed by induce express in E． coli BL2l ( DE3) ． The SDS-PAGE gel containing foreign protein was immunized BALB /c mice to prepare antisera． Indirect immunofluorescent assay ( IFA) and Western blot assay were performed． Ｒesult The mice anti-pET-chIL-10 specific serum could react with eukaryotic expressed chIL-10 in IFA and Western blot assay． Conclusion Antiserum to prokaryotic expressed chicken IL-10 could be used in vivo research study．

Key words: <p>Chicken interleukin-10, prokaryotic expression, mice antiserum, in vivo assay</p>

摘要： 摘要: 目的获得鸡白介素-10( chIL-10) 的原核表达产物，制备鼠抗血清，分析与真核系统表达的chIL-10 的反应原性。方法采用ＲT-PCＲ 从经脂多糖( LPS) 体外刺激的鸡外周血淋巴细胞中扩增编码成熟IL-10 的基因，克隆至原核表达载体pET-30a( + ) ，成功构建原核重组质粒pET-chIL-10; 转化至大肠杆菌DH5α 中IPTG 诱导表达，切下含有重组蛋白的SDS-PAGE 胶条，免疫制备鼠抗血清; 分别与包含信号肽和去除信号肽的chIL-10 真核表达细胞系CHO-euchIL-10F 和CHO-euchIL-10M 进行间接免疫荧光抗体试验( IFA) 和蛋白免疫印迹试验( Western blot) ，验证有无反应性。结果成功构建chIL-10 的原核表达载体pET-chIL-10，获得了高效价的鼠抗血清，能与表达chIL-10的真核细胞系反应。结论原核表达的chIL-10 特异性鼠抗血清将能用于体内研究的检测。

关键词: <, p>, 鸡白介素-10, 原核表达, 鼠抗血清, 体内实验<, /p>