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.
出处
《安徽农业科学》
CAS
北大核心
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
