甲型H1N1流感病毒HA1、NAe重组蛋白的表达及纯化效率研究

目的研究甲型H1N1流感病毒血凝素HA1及神经氨酸酶NAe重组蛋白的表达效率及纯化方法。方法在2009年广东省首株甲型H1N1流感病毒的HA和NA全长基因克隆的基础上,去除其信号肽（或信号锚定序列）和跨膜结构,在原核表达体系中表达HA1和NAe蛋白,采用SDS-PAGE和Western-blot法研究蛋白的表达效率、IPTG诱导作用及His Bind树脂纯化对蛋白免疫活性的影响。结果获得了具有免疫活性的高浓度和高纯度的HA1-57 290 Mr及NAe-69 280 Mr重组蛋白。结论截短优化的甲型H1N1流感病毒HA1、NAe重组基因经IPTG诱导可在原核表达体系中高效表达且纯化后具有较好的免疫活性。

Study on the efficiency of expression and purification of influenza A H1N1 virus recombinant proteins HA1 and NAe

Objectives To study the expression and purification efficiency of influenza A H1N1 virus recombinant proteins HA1 and NAe.Methods Recombinant proteins HA1 and NAe without signal peptide （or signal-anchor sequence）and transmembrane region were expressed in procaryon expression system based on recombinant plasmids containing HA and NA whole genes amplified from the first isolate of 2009 influenza A H1N1 virus in Guangdong province.The expression efficiency,impact of IPTG induction and His Bind resin purification,and immunological activity of the recombinant proteins were evaluated using SDS-PAGE and Western-blot.Results The HA1-57 290 Mr and NAe-69 280 Mr recombinant proteins with immunological activity were obtain by using procaryon expression system,IPTG induction and His Bind resin purification.Conclusion The recombinant genes HA1 and NAe could efficiently expressed in procaryon expression system with IPTG induction.And after purification,it did not damage their immunological activity.