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学科主题: Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology
题名: Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody
作者: Xiao, Nan; Zhang, Xiaohua; Dai, Linfeng; Yuan, Li; Wang, Yuzhen; Zhang, Min; Xu, Tao; Dai, Heping
通讯作者: Dai, HP, Chinese Acad Sci, Inst Hydrobiol, 7 So E Lake Rd, Wuhan 430072, Hubei, Peoples R China
关键词: white spot syndrome virus (WSSV) ; phage display ; scFvAl ; WSSV388
刊名: JOURNAL OF VIROLOGICAL METHODS
发表日期: 2006-11-01
DOI: 10.1016/j.jviromet.2006.06.027
卷: 137, 期:2, 页:272-279
收录类别: SCI
文章类型: Article
部门归属: Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China; Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
摘要: In a previous study, a scFv phage display library against white spot syndrome virus (WSSV) was constructed and yielded a clone designated A I with conformational specificity against native but not denatured viral antigen. Although the clone A1 has been used successfully as a diagnostic antibody, its precise target antigen has not been elucidated. A different strategy was adopted involving the construction of a second T7 phage display library utilizing mRNA isolated from shrimp infected with WSSV. Following RT-PCR and T7 phage library construction, phages displaying the candidate epitope were selected with A I scFv. Since successive enrichment steps were not associated with an increased titer of the phages, enrichment after successive tests was confirmed by PCR resulting in the prefer-red selection of a specific DNA sequence encoding a novel nucleocapsid protein WSSV388. Immune electron microscopy revealed that WSSV388 is located on the nucleocapsid. This result demonstrated that unknown antigen could be identified by phage display using the epitope conformation dependent scFv. (c) 2006 Elsevier B.V. All rights reserved.
英文摘要: In a previous study, a scFv phage display library against white spot syndrome virus (WSSV) was constructed and yielded a clone designated A I with conformational specificity against native but not denatured viral antigen. Although the clone A1 has been used successfully as a diagnostic antibody, its precise target antigen has not been elucidated. A different strategy was adopted involving the construction of a second T7 phage display library utilizing mRNA isolated from shrimp infected with WSSV. Following RT-PCR and T7 phage library construction, phages displaying the candidate epitope were selected with A I scFv. Since successive enrichment steps were not associated with an increased titer of the phages, enrichment after successive tests was confirmed by PCR resulting in the prefer-red selection of a specific DNA sequence encoding a novel nucleocapsid protein WSSV388. Immune electron microscopy revealed that WSSV388 is located on the nucleocapsid. This result demonstrated that unknown antigen could be identified by phage display using the epitope conformation dependent scFv. (c) 2006 Elsevier B.V. All rights reserved.
关键词[WOS]: SPOT SYNDROME VIRUS ; STRUCTURAL PROTEINS ; SHRIMP ; GENE ; AFFINITY ; CLONING ; GENOME ; CELLS
语种: 英语
WOS记录号: WOS:000241432400014
ISSN号: 0166-0934
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/8810
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Xiao, Nan; Zhang, Xiaohua; Dai, Linfeng; Yuan, Li; Wang, Yuzhen; Zhang, Min; Xu, Tao; Dai, Heping.Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody,JOURNAL OF VIROLOGICAL METHODS,2006,137(2):272-279
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