目的表达并纯化人甲型流感病毒H1N1亚型NS1蛋白。方法用RT-PCR法从人甲型流感病毒株A/PR/8/34(H1N1)中扩增NS1基因,克隆入原核表达载体pTXB1,构建重组原核表达质粒pTXB1/NS1,经酶切鉴定后,转化大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE分析表达形式和表达量。经几丁质柱亲和层析纯化表达蛋白,串联飞行时间质谱仪检测其相对分子质量。结果所构建的重组表达质粒pTXB1/NS1序列完整,插入的基因片段全长690bp。以1.0mmol/LIPTG37℃诱导4h,重组蛋白表达量最高,占菌体总蛋白的50%以上。破菌上清及沉淀中均有目的蛋白表达。纯化的NS1蛋白纯度达95%以上,相对分子质量约为26000。结论已成功表达并纯化了人甲型流感病毒H1N1亚型NS1蛋白,为其进一步的研究奠定了基础。 Objective To express and purify the H1N1 subtype NS1 protein of human influenza A virus. Methods The NS1 gene was amplified from human influenza A virus strain A / PR / 8/34 (H1N1) by RT-PCR and cloned into prokaryotic expression vector pTXB1 to construct recombinant prokaryotic expression plasmid pTXB1 / NS1. , Transformed into E. coli BL21 (DE3), induced by IPTG and analyzed by SDS-PAGE. Chitin column affinity chromatography purified protein, tandem time of flight mass spectrometry to detect the relative molecular mass. Results The constructed recombinant plasmid pTXB1 / NS1 was complete and the inserted gene was 690bp in length. Induced with 1.0mmol / L IPTG at 37 ℃ for 4h, the highest expression level of recombinant protein accounted for more than 50% of the total bacterial protein. Broken-bacteria supernatant and sediment in the target protein expression. Purified NS1 protein purity of more than 95%, the relative molecular mass of about 26,000. Conclusion The H1N1 subtype NS1 protein of human influenza A virus has been successfully expressed and purified, which lays the foundation for its further research.