| 奶牛PrPc结构蛋白多肽合成与免疫学特性分析 |
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| 引用本文： | 杨建民,赵德明,郝永新,王辉暖,李宁. 奶牛PrPc结构蛋白多肽合成与免疫学特性分析[J]. 中国兽医杂志, 2007, 43(1): 3-7 |
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| 作者姓名： | 杨建民 赵德明 郝永新 王辉暖 李宁 |
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| 作者单位： | 1. 中国农业大学国家动物海绵状脑病实验室,北京,海淀,100094
2. 中国农业大学农业生物技术国家重点实验室,北京,海淀,100094 |
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| 基金项目： | 国家自然科学基金;高等学校博士学科点专项科研项目 |
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| 摘 要： | 目的预测奶牛PrPc结构蛋白抗原表位,人工合成特定序列奶牛PrPc多肽用于抗体制备,同时为研究特定表位结构蛋白的功能特性提供有效手段。方法运用Goldenkey分子生物学软件对牛PrPc结构蛋白的抗原表位及其二级结构进行了分析比较,结合现有牛PrPc单克隆抗体识别表位从中筛选了一段显示表位特征的氨基酸残基序列,应用固相合成法合成15个氨基酸残基的奶牛PrPc多肽,并利用MBS法将其与KLH偶连制成免疫原,应用于动物免疫及多克隆抗体的制备,对该多抗与PrPc的反应性进行了检测。结果偶连后的牛PrPc多肽具有免疫原性,并获得了抗牛PrPc多克隆抗体。结论奶牛PrPc合成肽抗原的获得和应用,可以部分替代天然抗原,为进一步制备PrP单克隆抗体,以及Prion疾病的深入研究提供了有利条件,同时为研究特定表位结构蛋白的功能特性提供了新的途径。
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| 关 键 词： | 合成肽 免疫学特性 |
| 文章编号： | 0529-6005（2007）01-0003-04 |
| 修稿时间： | 2005-03-25 |
Synthesizing and immunization analysis of bovine PrPc structural protein peptides |
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| YANG Jian-min,ZHAO De-ming,HAO Yong-xin,WANG Hui-nuan,LI Ning. Synthesizing and immunization analysis of bovine PrPc structural protein peptides[J]. Chinese Journal of Veterinary Medicine, 2007, 43(1): 3-7 |
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| Authors: | YANG Jian-min ZHAO De-ming HAO Yong-xin WANG Hui-nuan LI Ning |
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| Affiliation: | 1. National Animal Transmissible Spongiform Eneephalopathies Laboratory, College of Veterinary Medicine,Beijing 100094, China; 2. State Key Laboratories of Agrobiology and Biotechnology, China Agricultural University, Beijing 100094, China |
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| Abstract: | Through forecasting the epitopes of bovine PrP~c structural protein and synthesis of peptides for special sequence, and antibody against prion could be prepared to offer effective methods for studying characters of PrP~c structural protein. The synthetic antigens were designed and conjugated with the keyhole limpet hemoyanin (KLH), according to the functional eptitopes of bovine prion, and the functional etitopes was determined by Golden Key software. rabbits were immunized with this antigen and the antibody was examined. The results showed that the antibody titer was as high as 1:10~8. Through Western blotting analysis, this peptide antibodies could react with both recombinant protein and purified bovine PrP~c. |
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| Keywords: | PrPc KLH |
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