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Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43
Mourez, Thomas1; Vabret, Astrid1; Han, Yang1; Dina, Julia1; Legrand, Loic1; Corbet, Sandrine1; Freymuth, Francois1
关键词Coronavirus Oc43 Strain Nucleocapsid Protein Baculovirus Insect Cells Western Blot
刊名JOURNAL OF VIROLOGICAL METHODS
2007-02-01
DOI10.1016/j.jviromet.2006.09.024
139期:2页:175-180
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
研究领域[WOS]Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
关键词[WOS]RECOMBINANT BACULOVIRUSES ; HUMAN CORONAVIRUS ; IDENTIFICATION ; INFECTION ; PNEUMONIA ; DIAGNOSIS ; ELISA ; GENE ; OC43
英文摘要

The nucleocapsid (N) gene of human coronavirus strain OC43 (HCoV-OC43) was amplified by reverse transcriptase-polymerase chain reaction, and cloned in pENTR (TM)/D-TOPO (R) plasmid. This plasmid containing the N gene was recombined with in a BaculoDirect (TM) baculovirus DNA designed in order to express N protein in fusion with a C-terminal polyhistidine tag containing V5 epitope. Sf21 cells were transfected with recombinant baculovirus DNA. Recombinant N protein was extracted from infected cells, analysed by SDS-PAGE and Western blot, and purified by Ni2+ affinity procedure. Sera from 100 healthcare workers and five 2-3-year-old children were tested in a Western blot assay using the purified recombinant N protein. All of the sera from adults and two of the sera from children have a positive result. (c) 2006 Elsevier B.V. All rights reserved.

语种英语
WOS记录号WOS:000244090300007
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被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/52079
专题北京大学第一临床医学院_感染疾病科
作者单位1.Univ Hosp Caen, Lab Human & Mol Virol, F-14033 Caen, France
2.Peking Univ, Med Coll Hosp, Dept Infect Dis, Beijing 100871, Peoples R China
推荐引用方式
GB/T 7714
Mourez, Thomas,Vabret, Astrid,Han, Yang,et al. Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43[J]. JOURNAL OF VIROLOGICAL METHODS,2007,139(2):175-180.
APA Mourez, Thomas.,Vabret, Astrid.,Han, Yang.,Dina, Julia.,Legrand, Loic.,...&Freymuth, Francois.(2007).Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43.JOURNAL OF VIROLOGICAL METHODS,139(2),175-180.
MLA Mourez, Thomas,et al."Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43".JOURNAL OF VIROLOGICAL METHODS 139.2(2007):175-180.
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