小反刍兽疫病毒H基因在杆状病毒中的表达及其免疫原性研究

doi:10.11843/j.issn.0366-6964.2014.06.016

Abstract

Abstract:

The aim of this study was to study the expression and immunogenicity of H protein of peste des petits ruminants virus (PPRV).The H gene of PPRV Nigeria75/1 strain was amplified by RT-PCR，and cloned into the donor plasmid pFB-LIC-Bse.After being sequenced，the recombinant plasmid pFB-LIC-Bse-PPRV-H was transformed into competent cells of E.coli DH10Bac to get recombinant shuttle plasmid.The recombinant shuttle plasmid was then transfected into sf21 cells to get recombinant baculovirus.Western blot，IFA and immunity tests in mice were performed to identify the immunoreactivity and immunogenicity of the expression products.The recombinant H protein with molecular mass of approximately 68 kDa was detected with His tag monoclonal antibody and PPRV positive serum.The exprsssion products possessed a favorable immunogenicity and fall immunized mice could produce anti-PPRV neutralizing antibody.The cell immune responses were examined by lymphocyte proliferative assay using MTS method.The results showed that the rBacmid-H was able to express H proteins in Vero cells.Furthermore，specific antibodies were induced in the second week after primary vaccination.In conclusion，the recombinant H protein had good immunogenicity and immunoreactivity，which laid a foundation of developing a new vaccine against PPR.

Key words: peste des petits ruminants virus, H protein, baculovirus expression system, immunogenicity detecting

CLC Number:

S852.659.5

SUI Xiu-kun，JIN Hong-yan，LI Wen-chao，SHI Li-jun，ZHAO Zhan-zhong，LIANG Lin，LI Gang. Expression of H Gene of Peste des Petits Ruminants Virus in Baculovirus and Its Immunogenicity Study[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(6): 974-980.

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Expression of H Gene of Peste des Petits Ruminants Virus in Baculovirus and Its Immunogenicity Study

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