鸡传染性支气管炎病毒HH06株囊膜蛋白多克隆抗体制备及生物学功能检测

doi:10.16431/j.cnki.1671-7236.2016.08.006

Abstract

Abstract:

In this study,IBV HH06 complete E gene was firstly cloned and sequenced.According to the hydrophilicity and antigenic index analysis,its partial gene (193 to 327 bp) was subcloned into prokaryotic expression vector pET-32a(+) and eukaryotic expression vector PVAX1.The recombinant plasmid pET-32a-E1 was transformed into E.coli Rosetta (DE3) and induced with IPTG.The recombinant IBV truncated E1 protein with molecular weight of 23 ku was observed as expected.It could be recognized by positive IBV antisera in Western blotting with high reactivity.Then the purified recombinant protein was used as antigen for immunization of rabbit to prepare polyclonal antibody.Indirect ELISA showed that the titer of polyclonal antibody was 2²⁰,and it had high reactivity and specialty with recombinant protein.Furthermore,IFA test demonstrated that this polyclonal antibody could react with Hela cells transfected with PVAX-E1 plasmid and IBV-infected CEK cells.The IBV E polyclonal antibody obtained in this study laid a foundation for further functional research of E protein in IBV pathogenesis.

Key words: IBV; envelope protein; polyclonal antibody; Western blotting; indirect fluorescence antibody assay

CLC Number:

S852.65

BAI Yan, ZHAO Lei, DU Wei-wei, PAN long, HUANG Xiao-dan, YANG Gui-jun, LI Guang-xing. Preparation of Polyclonal Antibody against Recombinant E Protein of IBV HH06 Strain and Its Biological Function[J]. , 2016, 43(8): 1967-1974.

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Preparation of Polyclonal Antibody against Recombinant E Protein of IBV HH06 Strain and Its Biological Function

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