The C-terminal portion of the nucleocapsid protein demonstrates SARS-CoV antigenicity.
In order to develop clinical diagnostic tools for rapid detection of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.
Liu
Guozhen
G
College of Life Sciences, Hebei Agricultural University, Baoding 071001, China.
Hu
Shaohui
S
Hu
Yongwu
Y
Chen
Peng
P
Yin
Jianning
J
Wen
Jie
J
Wang
Jingqiang
J
Lin
Liang
L
Liu
Jinxiu
J
You
Bo
B
Yin
Ye
Y
Li
Shuting
S
Wang
Hao
H
Ren
Yan
Y
Ji
Jia
J
Zhao
Xiaoqian
X
Sun
Yongqiao
Y
Zhang
Xiaowei
X
Fang
Jianqiu
J
Wang
Jian
J
Liu
Siqi
S
Yu
Jun
J
Zhu
Heng
H
Yang
Huanming
H
D000956:Antigens, Viral / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D016679:Genome, Viral; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000276:immunology; D015003:Yeasts / Q000235:genetics
D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins