Recombinant hepatitis B virus core particles: Association, dissociation and encapsidation of green fluorescent protein
Recombinant hepatitis B virus core particles: Association, dissociation and encapsidation of green fluorescent protein
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DOI：
10.1016/j.jviromet.2008.05.025
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发表时间：
2008-08-01
影响因子：
3.1
通讯作者：
Tan, Wen Siang
中科院分区：
文献类型：
--
作者：
Lee, Khai Wooi;Tan, Wen Siang
在大肠杆菌中表达的重组乙型肝炎病毒（HBV）核心抗原（HBcAg）可自组装成约35 nm的二十面体衣壳，可用作基因或药物递送载体。研究了 C 端截短的 HBcAg 与尿素和盐酸胍 (GdnHCl) 的结合和解离特性。透射电子显微镜 (TEM) 显示，当物理去除所施加的变性剂时，解离的 HBcAg 能够重新结合成颗粒。为了评估颗粒捕获分子的潜力，将纯化的绿色荧光蛋白 (GFP) 应用于解离的 HBcAg 进行衣壳化。使用蔗糖密度梯度超速离心纯化含有 GFP 分子的 HBcAg 颗粒，并使用天然琼脂糖凝胶电泳和 TEM 进行分析。建立了一种将 GFP 包裹在 HBcAg 颗粒中的方法，该方法具有捕获药物或核酸的潜力。 (c) 2008 Elsevier B.V. 保留所有权利。
The recombinant hepatitis B virus (HBV) core antigen (HBcAg) expressed in Escherichia coli self-assembles into icosahedral capsids of about 35 nm which can be exploited as gene or drug delivery vehicles. The association and dissociation properties of the C-terminally truncated HBcAg with urea and guanidine hydrochloride (GdnHCl) were Studied. Transmission electron microscopy (TEM) revealed that the dissociated HBcAg was able to re-associate into particles when the applied denaturing agents were physically removed. In order to evaluate the potential of the Particles in capturing molecules, purified green fluorescent protein (GFP) was applied to the dissociated HBcAg for encapsidation. The HBcAg particles harbouring the GFP molecules were Purified using sucrose density gradient ultracentrifugation and analysed using native agarose gel electrophoresis and TEM. A method for the encapsidation of GFP in HBcAg particles which has the potential to Capture drugs or nucleic acids was established. (c) 2008 Elsevier B.V. All rights reserved.