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Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody
Xiao, Nan; Zhang, Xiaohua; Dai, Linfeng; Yuan, Li; Wang, Yuzhen; Zhang, Min; Xu, Tao; Dai, Heping; Dai, HP, Chinese Acad Sci, Inst Hydrobiol, 7 So E Lake Rd, Wuhan 430072, Hubei, Peoples R China
2006-11-01
Source PublicationJOURNAL OF VIROLOGICAL METHODS
ISSN0166-0934
Volume137Issue:2Pages:272-279
AbstractIn 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.; 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.
SubtypeArticle
KeywordWhite Spot Syndrome Virus (Wssv) Phage Display Scfval Wssv388
DepartmentChinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China; Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaBiochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
DOI10.1016/j.jviromet.2006.06.027
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
WOS SubjectBiochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
WOS IDWOS:000241432400014
WOS KeywordSPOT SYNDROME VIRUS ; STRUCTURAL PROTEINS ; SHRIMP ; GENE ; AFFINITY ; CLONING ; GENOME ; CELLS
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8810
Collection期刊论文
Corresponding AuthorDai, HP, Chinese Acad Sci, Inst Hydrobiol, 7 So E Lake Rd, Wuhan 430072, Hubei, Peoples R China
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Xiao, Nan,Zhang, Xiaohua,Dai, Linfeng,et al. Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody[J]. JOURNAL OF VIROLOGICAL METHODS,2006,137(2):272-279.
APA Xiao, Nan.,Zhang, Xiaohua.,Dai, Linfeng.,Yuan, Li.,Wang, Yuzhen.,...&Dai, HP, Chinese Acad Sci, Inst Hydrobiol, 7 So E Lake Rd, Wuhan 430072, Hubei, Peoples R China.(2006).Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody.JOURNAL OF VIROLOGICAL METHODS,137(2),272-279.
MLA Xiao, Nan,et al."Isolation and identification of a novel WSSV nucleocapsid protein by cDNA phage display using an scFv antibody".JOURNAL OF VIROLOGICAL METHODS 137.2(2006):272-279.
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