expression, purification and glycosylation analysis of chicken infectious bursal disease virus vp2 in yeast
نویسندگان
چکیده
infectious bursal disease (ibd), a highly contagious and devastating disease in young chicken, is causedby infectious bursal disease virus (ibdv). to improve the immunogenicity of recombinant ibdv subunitvaccine, an attempt was made to find a new way to prepare ibd vaccine containing glycosylated mvp2antigen. firstly, ibdv mvp2 gene (with a nucleic acid sequence encoding b cell epitope of ibdv(kfdqml) in the 5′-end of the vp2, with a nucleic acid sequence encoding b cell epitope of ibdv (lasp)and (his) 6-tag in the 3′-end of the vp2) was cloned. secondly, ibdv mvp2 protein was expressed in themethylotrophic yeast pichia pastoris which can secret glycosylated protein. the recombinant mvp2 proteincould be stained pink with periodic acid-schiff reagents (pas), which showed that mvp2 was glycosylated.finally, ibdv mvp2 protein was purified with his-trap (1 ml) affinity chromatography. these resultsindicate that glycosylated ibdv vp2 protein modified with epitope peptides can be expressed in pichiapastoris, which lay the groundwork for the development of a recombinant infectious bursal disease vaccinewith high immunogenicity.
منابع مشابه
Expression, purification and glycosylation analysis of chicken infectious bursal disease virus VP2 in yeast
Infectious bursal disease (IBD), a highly contagious and devastating disease in young chicken, is causedby infectious bursal disease virus (IBDV). To improve the immunogenicity of recombinant IBDV subunitvaccine, an attempt was made to find a new way to prepare IBD vaccine containing glycosylated mVP2antigen. Firstly, IBDV mVP2 gene (with a nucleic acid sequence encoding B cell epitope of IBDV(...
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Infectious bursal disease (IBD) is an economically important viral disease of chickens with worldwide distribution which suppresses the immune system of young chickens. VP2 is the major host-protective protein of infectious bursal disease virus (IBDV). The encoding region of VP2 protein was PCR amplified from a plasmid containing a cDNA fragment of large genomic segment of IBDV, strain D78. Thi...
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Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. The expression of heterologous proteins in P.pastoris is fast, simple and inexpensive. In this study, VP2 encoding gene of classical D78 IBDV was amplified using reverse transcription (RT) polymerase chain reaction (PCR) and cloned into pPICZαA vector...
متن کاملcloning and secretory expression of vp2 gene of infectious bursal disease virus in eukaryotic cells
vp2 gene coding region of a vaccinal strain (d78) of infectious bursal disease virus (ibdv) was clonedin a eukaryotic expression vector, psec tag2a. the gene was placed downstream of ig κ chain leadersequence, under the control of human cytomegalovirus (hcmv) immediate early enhancer and promoter. theconstruct psec tag2a-vp2 was transfected in cos-7 cell line and the expression and secretion of...
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عنوان ژورنال:
iranian journal of veterinary researchناشر: shiraz university
ISSN 1728-1997
دوره 14
شماره 3 2013
کلمات کلیدی
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