Construction of cell line stably expressing IBDV VP 2 protein mutants

A couple of primers was designed on the base of VP2 gene sequence of infectious bursal disease virus (IBDV) HZ2 strain, and the VP2 gene was successfully cloned into eukaryotic expression vector pDsRed2-N1. Then, the recombinant plasmid pDsRed2-N1-VP2 was constructed. The 279 site and 284 site on VP...

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Main Authors: SUN Jing, WEI Yong-wei, ZHANG Xiao-dong, YU Lian
Format: Article
Language:English
Published: Zhejiang University Press 2011-09-01
Series:浙江大学学报. 农业与生命科学版
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Online Access:https://www.academax.com/doi/10.3785/j.issn.1008-9209.2011.05.001
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author SUN Jing
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
author_facet SUN Jing
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
author_sort SUN Jing
collection DOAJ
description A couple of primers was designed on the base of VP2 gene sequence of infectious bursal disease virus (IBDV) HZ2 strain, and the VP2 gene was successfully cloned into eukaryotic expression vector pDsRed2-N1. Then, the recombinant plasmid pDsRed2-N1-VP2 was constructed. The 279 site and 284 site on VP2 protein were mutated singly or homeochronously through site-directed mutagenesis on the constructed vector pDsRed2-N1-VP2. By this way, three mutants of P279, P284 and PDA were constructed in next step. The Vero cell line stably expressing VP2 protein and this mutant were also constructed successfully on the base of P279, P284 and PDA by indirect immunofluorescence test. The recombination plasmid and cell line constructed lay the foundation on the deep research about the role and mechanism of 279 site and 284 site in virulence change and antigenic variation of IBDV, which also play a significant role in exploring more efficient and cheaper IBDV DNA vaccination.
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publisher Zhejiang University Press
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series 浙江大学学报. 农业与生命科学版
spelling doaj-art-031596173de5459c8c8d159c1a89a8a12025-08-20T03:16:03ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552011-09-013747347810.3785/j.issn.1008-9209.2011.05.00110089209Construction of cell line stably expressing IBDV VP 2 protein mutantsSUN JingWEI Yong-weiZHANG Xiao-dongYU LianA couple of primers was designed on the base of VP2 gene sequence of infectious bursal disease virus (IBDV) HZ2 strain, and the VP2 gene was successfully cloned into eukaryotic expression vector pDsRed2-N1. Then, the recombinant plasmid pDsRed2-N1-VP2 was constructed. The 279 site and 284 site on VP2 protein were mutated singly or homeochronously through site-directed mutagenesis on the constructed vector pDsRed2-N1-VP2. By this way, three mutants of P279, P284 and PDA were constructed in next step. The Vero cell line stably expressing VP2 protein and this mutant were also constructed successfully on the base of P279, P284 and PDA by indirect immunofluorescence test. The recombination plasmid and cell line constructed lay the foundation on the deep research about the role and mechanism of 279 site and 284 site in virulence change and antigenic variation of IBDV, which also play a significant role in exploring more efficient and cheaper IBDV DNA vaccination.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2011.05.001infectious bursal disease virusVP2 genesite-directed mutagenesismutantVero cell line
spellingShingle SUN Jing
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
Construction of cell line stably expressing IBDV VP 2 protein mutants
浙江大学学报. 农业与生命科学版
infectious bursal disease virus
VP2 gene
site-directed mutagenesis
mutant
Vero cell line
title Construction of cell line stably expressing IBDV VP 2 protein mutants
title_full Construction of cell line stably expressing IBDV VP 2 protein mutants
title_fullStr Construction of cell line stably expressing IBDV VP 2 protein mutants
title_full_unstemmed Construction of cell line stably expressing IBDV VP 2 protein mutants
title_short Construction of cell line stably expressing IBDV VP 2 protein mutants
title_sort construction of cell line stably expressing ibdv vp 2 protein mutants
topic infectious bursal disease virus
VP2 gene
site-directed mutagenesis
mutant
Vero cell line
url https://www.academax.com/doi/10.3785/j.issn.1008-9209.2011.05.001
work_keys_str_mv AT sunjing constructionofcelllinestablyexpressingibdvvp2proteinmutants
AT weiyongwei constructionofcelllinestablyexpressingibdvvp2proteinmutants
AT zhangxiaodong constructionofcelllinestablyexpressingibdvvp2proteinmutants
AT yulian constructionofcelllinestablyexpressingibdvvp2proteinmutants