| SARS-CoVN蛋白抗原表位的筛选和鉴定 |
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| 引用本文： | 黄欣 高亚萍 王希良 辛忠涛 董洁 邵宁生 柳川. SARS-CoVN蛋白抗原表位的筛选和鉴定[J]. Virologica Sinica, 2006, 21(4): 314-318 |
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| 作者姓名： | 黄欣 高亚萍 王希良 辛忠涛 董洁 邵宁生 柳川 |
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| 作者单位： | 军事医学科学院基础医学研究所 北京100850(黄欣，高亚萍，辛忠涛，董洁，邵宁生，柳川)，军事医学科学院微生物流行病研究所 北京100071(王希良) |
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| 基金项目： | 国家自然科学基金资助项目(30340024) |
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| 摘 要： | 利用噬菌体展示的线性12肽库从马抗SARS-CoVIgG筛选SARS-CoV的抗原表位。经生物淘洗富集的噬菌体克隆被测序。获得两个共有序列:DXXDP和TXTLL。它们分别与SARS-CoVN蛋白341-345和392-396位氨基酸序列高度同源。含共有序列的克隆在ELISA竞争抑制试验中与SARS-CoVN蛋白竞争结合马抗SARS-CoVIgG。将两个共有序列肽通过基因重组技术成功展示到大肠杆菌鞭毛,获得重组菌F1和F2。用重组菌F1和F2免疫接种试验Balb/c小鼠产生的血清均能与SARS-CoVN蛋白特异结合。说明DXXDP和TXTLL是SARS-CoVN蛋白的两个连续抗原表位。
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| 关 键 词： | SARS-CoV N蛋白 噬菌体展示 抗原表位 |
| 收稿时间： | 2005-12-05 |
| 修稿时间： | 2006-01-10 |
Screening and Identification of the SARS-CoV N Protein Epitopes |
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| HUANG Xin, GAO Ya-ping, WANG Xi-liang, XIN Zhong-tao, DONG Jie, SHAO Ning-sheng, LIU Chuan. Screening and Identification of the SARS-CoV N Protein Epitopes[J]. 中国病毒学(英文版), 2006, 21(4): 314-318 |
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| Authors: | HUANG Xin GAO Ya-ping WANG Xi-liang XIN Zhong-tao DONG Jie SHAO Ning-sheng LIU Chuan |
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| Affiliation: | 1.Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China; 2. Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China |
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| Abstract: | The epitopes of SARS-CoV were screened from a 12-mer phage display random peptide library using anti- SARS-CoV horse polyclonal antibodies of as a target. The phage clones enriched in biopannings were sequenced. Two consensus sequences were obtained, DXXDP and TXTLL, which had close identity to the 341-345 aa and 392-396 aa of SARA-CoV N protein sequences,respectively. The phage clones with consensus sequences and the N protein were both recognized and bound by the antibodies in a competitive-inhibition ELISA test. The consensus sequence peptides were cloned and displayed on the bacterial flagellin. Balb/c mice were vaccinated using the reconstruted bacteria and the collected serum was shown to speccifically combined with SARS-CoV N protein. This confirmed that DXXDP and TXTLL are two continuous epitopes of SARS-CoV N protein. |
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| Keywords: | SARS-CoV N protein Phage display Epitope |
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