CAJ | 학술논문

[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 F_(2-3))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NDV F_(2-3)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 bp ChIFN-γ与NDV F_(2-3)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NDV F_(2-3)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.
[목적]연구계γ-간우소(ChIFN-γ)적면역좌제공능,구건ChIFN-γ여신성역 F단백항원표위(NDV F_(2-3))적융합기인,병진행원핵표체.[방법]이두미상련적방식구건계γ-간우소기인여NDV F_(2-3)적pET-32a중조질립,경PCR확증、쌍매절화측서감정후,중조자재E.coliBL21세포진행IPTG유도융합단백표체,채용SDS-PAGE화Western blot방법분별검측표체적산물.[결과]획득료726 bp ChIFN-γ여NDV F_(2-3)융합기인,경원핵표체,융합단백분자량약위35 000,능여상응적항체결합.[결론]ChIFN-γ여NDV F_(2-3)천련기인재원핵세포중능유효표체,차융합단백구유일정적면역활성.
[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.