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

[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。

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

[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 F_(2-3)) was built and expressed in prokaryotic cells. [Method] Using enzyme digestion, chicken interferon-γ was inserted into pET-32a-NDV F_(2-3) 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 F_(2-3) could effectively express in prokaryotic cells and had a certain immune activity.