Screening and Identification of the SARS-CoV N Protein Epitopes

HUANG Xin; GAO Ya-ping; WANG Xi-liang; XIN Zhong-tao; DONG Jie; SHAO Ning-sheng; LIU Chuan

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August 2006

HUANG Xin, GAO Ya-ping, WANG Xi-liang, XIN Zhong-tao, DONG Jie, SHAO Ning-sheng and LIU Chuan. Screening and Identification of the SARS-CoV N Protein Epitopes[J]. Virologica Sinica, 2006, 21(4): 314-318.

Citation: 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 .VIROLOGICA SINICA, 2006, 21(4) : 314-318.

SARS-CoV N蛋白抗原表位的筛选和鉴定

黄欣 ¹ ,
高亚萍 ¹ ,
王希良 ² ,
辛忠涛 ¹ ,
董洁 ¹ ,
邵宁生 ¹ ,
柳川 ^1*

1.
1.军事医学科学院基础医学研究所，北京100850
2.
军事医学科学院微生物流行病研究所，北京100071

摘要

利用噬菌体展示的线性12肽库从马抗SARS-CoV IgG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-CoV N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-CoV N蛋白竞争结合马抗SARS-CoV IgG。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-CoV N蛋白特异结合。说明DXXDP和TXTLL是SARS-CoV N蛋白的两个连续抗原表位。
- SARS-CoV
- , N蛋白
- , 噬菌体展示
- , 抗原表位

Screening and Identification of the SARS-CoV N Protein Epitopes

1.
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

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.
- SARS-CoV
- , N protein
- , Phage display
- , Epitope

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Screening and Identification of the SARS-CoV N Protein Epitopes

1. 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

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.