Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells

In order to improve the immunity effect of infectious bursal disease virus (IBDV) polyprotein (VP2/4/3) DNA vaccine, fusion genes of IBDV polyprotein (VP2/4/3) gene and chicken interleukin 18 (ChIL-18) gene were constructed using the technique of splicing by overlapping extension (SOEing) and three...

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Main Authors: HOU Xian-hong, WEI Yong-wei, ZHANG Xiao-dong, YU Lian
Format: Article
Language:English
Published: Zhejiang University Press 2010-01-01
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.1008-9209.2010.01.001
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author HOU Xian-hong
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
author_facet HOU Xian-hong
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
author_sort HOU Xian-hong
collection DOAJ
description In order to improve the immunity effect of infectious bursal disease virus (IBDV) polyprotein (VP2/4/3) DNA vaccine, fusion genes of IBDV polyprotein (VP2/4/3) gene and chicken interleukin 18 (ChIL-18) gene were constructed using the technique of splicing by overlapping extension (SOEing) and three times PCR. These fusion gene fragments were directly cloned into pCI, an eukaryotic expression vector, to produce recombinant expression plasmids pCI-VP2/4/3-ChIL-18 and pCI-ChIL-18-VP2/4/3. Then, co-expressing plasmid co-pCI-VP2/4/3-ChIL-18 was constructed on the basis of pCI-VP2/4/3 and pCI-ChIL-18 constructed in our lab by PCR. These recombination plasmids were transfected into Vero cells introduced by Lipofectamine 2000 reagent. The interest proteins were detected by indirect immunofluorescence, indicating that these fusion genes were correctly expressed in Vero cells. It is provided a basis for the research of immune-enhancement of ChIL-18 and manufacturing highly effective and inexpensive DNA vaccination against IBDV.
format Article
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2097-5155
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publishDate 2010-01-01
publisher Zhejiang University Press
record_format Article
series 浙江大学学报. 农业与生命科学版
spelling doaj-art-3411c784ae114de5b098fb83d23a33bf2025-08-20T03:34:22ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552010-01-01361810.3785/j.issn.1008-9209.2010.01.00110089209Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cellsHOU Xian-hongWEI Yong-weiZHANG Xiao-dongYU LianIn order to improve the immunity effect of infectious bursal disease virus (IBDV) polyprotein (VP2/4/3) DNA vaccine, fusion genes of IBDV polyprotein (VP2/4/3) gene and chicken interleukin 18 (ChIL-18) gene were constructed using the technique of splicing by overlapping extension (SOEing) and three times PCR. These fusion gene fragments were directly cloned into pCI, an eukaryotic expression vector, to produce recombinant expression plasmids pCI-VP2/4/3-ChIL-18 and pCI-ChIL-18-VP2/4/3. Then, co-expressing plasmid co-pCI-VP2/4/3-ChIL-18 was constructed on the basis of pCI-VP2/4/3 and pCI-ChIL-18 constructed in our lab by PCR. These recombination plasmids were transfected into Vero cells introduced by Lipofectamine 2000 reagent. The interest proteins were detected by indirect immunofluorescence, indicating that these fusion genes were correctly expressed in Vero cells. It is provided a basis for the research of immune-enhancement of ChIL-18 and manufacturing highly effective and inexpensive DNA vaccination against IBDV.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.01.001infectious bursal disease virusgene fusionco-expressionDNA vaccineindirect immunofluorescence
spellingShingle HOU Xian-hong
WEI Yong-wei
ZHANG Xiao-dong
YU Lian
Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
浙江大学学报. 农业与生命科学版
infectious bursal disease virus
gene fusion
co-expression
DNA vaccine
indirect immunofluorescence
title Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
title_full Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
title_fullStr Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
title_full_unstemmed Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
title_short Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
title_sort construction of plasmids containing ibdv vp2 4 3 and chil 18 fusion genes and co expression genes as well as proteins expressed on vero cells
topic infectious bursal disease virus
gene fusion
co-expression
DNA vaccine
indirect immunofluorescence
url https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.01.001
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AT yulian constructionofplasmidscontainingibdvvp243andchil18fusiongenesandcoexpressiongenesaswellasproteinsexpressedonverocells