恒河猴干扰素-γ基因的原核表达及其免疫活性的初步研究

发布时间：2018-03-01 21:06

本文关键词： 恒河猴干扰素-γ 原核表达生物学活性　出处：《中国兽医科学》2017年03期 　论文类型：期刊论文

【摘要】：为构建以恒河猴干扰素-γ为目的基因的原核表达重组质粒并进行免疫活性分析,获得具有生物学活性重组IFN-γ蛋白。应用逆转录-聚合酶链式反应(RT-PCR)从恒河猴外周血单个核细胞(PBM C)扩增干扰素-γ基因,然后将其克隆至表达载体p ET32a(+)中,转化BL21(D E3)进行诱导表达。SDS-PAG E电泳分析其可溶性,W estern-blot检测纯化后的重组蛋白,采用淋巴细胞增殖试验检测重组IFN-γ的活性。结果显示,成功构建恒河猴干扰素-γ原核表达重组菌,重组蛋白分子质量约为35.4 ku,主要以包涵体形式存在。重组IFN-γ具有促进淋巴细胞转化增殖的生物学活性。结果表明,本试验成功获得了重组IFN-γ蛋白,表达量高且具有生物学活性,为重组IFN-γ治疗恒河猴疾病奠定了基础。
[Abstract]:In order to construct the prokaryotic expression plasmid of rhesus monkey interferon-纬 gene and analyze its immunological activity, The recombinant IFN- 纬 protein with biological activity was obtained. The IFN- 纬 gene was amplified from the peripheral blood mononuclear cells of rhesus monkey by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into the expression vector pET32a (). The recombinant protein was detected by soluble W estern-blot and the activity of recombinant IFN- 纬 was detected by lymphocyte proliferation test. The results showed that rhesus monkey IFN- 纬 prokaryotic expression strain was successfully constructed. The molecular weight of recombinant IFN- 纬 protein is about 35.4 ku.Recombinant IFN- 纬 has the biological activity of promoting lymphocyte transformation and proliferation. The results show that the recombinant IFN- 纬 protein has high expression and biological activity. It lays a foundation for the treatment of rhesus monkey disease by recombinant IFN- 纬.
【作者单位】：四川农业大学动物医学院;成都军区疾病预防控制中心;
【基金】：公益性行业(农业)科研专项(201303042) 国家自然科学基金项目(31272620) 国家重点研究发展计划项目(2016YFD0501009)
【分类号】：S858.9

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