鸡γ-干扰素与新城疫病毒F蛋白抗原表位融合基因的构建与表达  

Fusion Expression and Construction of Antigen Epitopes from F Gene of Newcastle Disease Virus with Chicken Interferon-gamma

作　　者：刘雪兰[1] 戴银[1] 程宝艳[2] 彭明义[1] 王骏俊[1] 余为一[1] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]安徽省农业科学院,安徽合肥230031

出　　处：《安徽农业科学》2010年第7期3862-3864,共3页Journal of Anhui Agricultural Sciences

基　　金：安农大稳定和引进人才资助;校长青年基金项目

摘　　要：[目的]研究鸡γ-干扰素(ChIFN-γ)的免疫佐剂功能,构建ChIFN-γ与新城疫F蛋白抗原表位(NDV F2-3)的融合基因,并进行原核表达。[方法]以头尾相连的方式构建鸡γ-干扰素基因与NDVF2-3的pET-32a重组质粒,经PCR扩增、双酶切和测序鉴定后,重组子在E.coliBL21细胞进行IPTG诱导融合蛋白表达,采用SDS-PAGE和Western blot方法分别检测表达的产物。[结果]获得了726 bpChIFN-γ与NDVF2-3融合基因,经原核表达,融合蛋白分子量约为35 000,能与相应的抗体结合。[结论]ChIFN-γ与NDVF2-3串联基因在原核细胞中能有效表达,且融合蛋白具有一定的免疫活性。[Objective] The aim was to study the immune adjuvant effects of chicken interferon-γ（ChIFN-γ）,a recombinant plasmid with ChIFN-γ and two antigen epitopes from F gene of Newcastle disease virus（NDV F2-3） was built and expressed in prokaryotic cells.[Method] Using enzyme digestion,chicken interferon-γ was inserted into pET-32a-NDV F2-3 expression vector,and then the recombinant pET-IFN-F was constructed and confirmed with PCR amplification,double restriction digestion and DNA sequencing transformed.The recombinant plasmid was expressed in E.coli BL21though IPTG.SDS-PAGE and Western blot were used to detect expression products.[Result] The length of recombinant gene was 726 bp by DNA sequencing.The fusion protein was about 35 000 and had the reactinogenicity with specific antibody.[Conclusion] The fusion gene encoding ChIFN-γ-NDV F2-3 could effectively express in prokaryotic cells and had a certain immune activity.

关 键 词：鸡IFN-γ NDVF2-3 融合蛋白 

分 类 号：S852.65[农业科学—基础兽医学]