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Prokaryotic expression and purification of HA1 and HA2 polypeptides for serological analysis of the 2009 pandemic H1N1 influenza virus

机译:用于2009大流行H1N1流感病毒血清学分析的HA1和HA2多肽的原核表达和纯化

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摘要

Hemagglutinin (HA) is an important influenza virus surface antigen that is highly topical in influenza research. In the present study, the genes encoding the HA1 and HA2 proteins from the 2009 pandemic influenza virus H1N1 (A/California/04/2009(H1N1)) were cloned into a prokaryotic expression plasmid pCold-TF, and soluble fusion proteins containing H1N1 HA1 and HA2 were produced by transformed Escherichia coil. Western blot assays were used to examine the immunoreactivity of the recombinant proteins using polyclonal and monoclonal antibodies derived against the whole virus A/California/04/2009(H1N1). Recombinant protein immunoreactivity was also analyzed qualitatively by ELISA and hemagglutination inhibition using human serum samples. These results will aid future immunological and serological studies of the 2009 pandemic H1N1 virus HA
机译:血凝素(HA)是一种重要的流感病毒表面抗原,在流感研究中非常重要。在本研究中,将来自2009年大流行性流感病毒H1N1(A / California / 04/2009(H1N1))的编码HA1和HA2蛋白的基因克隆到原核表达质粒pCold-TF中,并将含有H1N1 HA1的可溶性融合蛋白克隆HA2和HA2是通过转化的大肠杆菌产生的。使用蛋白质印迹法检测重组蛋白的免疫反应性,使用的是针对整个病毒A / California / 04/2009(H1N1)的多克隆和单克隆抗体。还使用人血清样品通过ELISA和血凝抑制作用对定性蛋白的免疫反应性进行了定性分析。这些结果将有助于将来对2009年大流行H1N1病毒HA的免疫学和血清学研究

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