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首页> 外文期刊>BMC research notes >Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris
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Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris

机译:使用甲基嗜热酵母毕赤酵母快速表达和纯化肝炎三角洲病毒抗原

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ObjectivePatients with dual hepatitis B (HBV) and hepatitis D (HDV) virus infection are at an increased risk of progression to liver cirrhosis and hepatocellular carcinoma than patients with a single viral infection. Treatment of viral hepatitis due to dual HBV/HDV infection represents a challenge. Currently there is no vaccine against HDV. Recombinant production of HDV antigen (HDAg) is the first step towards a potential vaccine candidate and the development of assays for HDV detection. ResultsThis study demonstrates the expression of one HDAg isoform, S-HDAg, in Pichia pastoris . A recombinant vector carrying a tagged gene encoding S-HDAg under the control of the methanol-inducible promoter AOX1 was designed and integrated into P. pastoris X33. The protein, which was purified using a Ni2+ affinity column and eluted at 100–150?mM imidazole, has potential as a recombinant antigen for further study.
机译:目的与双重病毒感染的患者相比,双重感染乙型肝炎(HBV)和丙型肝炎(HDV)的患者罹患肝硬化和肝细胞癌的风险更高。由双重HBV / HDV感染引起的病毒性肝炎的治疗是一个挑战。当前没有针对HDV的疫苗。 HDV抗原(HDAg)的重组生产是迈向潜在疫苗候选者和开发HDV检测方法的第一步。结果这项研究证明了一种HDAg亚型S-HDAg在毕赤酵母中的表达。设计了在甲醇诱导型启动子AOX1的控制下携带编码S-HDAg的标记基因的重组载体,并将其整合到巴斯德毕赤酵母X33中。该蛋白使用Ni 2 + 亲和柱纯化,并在100–150?mM咪唑中洗脱,具有作为重组抗原的潜力,有待进一步研究。

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