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人类 JC 病毒样颗粒包装的 DNA 大小分析。

Analysis of the size of DNA packaged by the human JC virus-like particle.

机构信息

Department of Medical Research, Chiayi Christian Hospital, Chiayi 60002, Taiwan.

出版信息

J Virol Methods. 2012 Jun;182(1-2):87-92. doi: 10.1016/j.jviromet.2012.03.017. Epub 2012 Mar 23.

Abstract

Previously, it has been demonstrated that the JC virus-like particle (VLP) is able to package DNA in E. coli and deliver the DNA into human colon cancer cells for gene expression. In this study, the maximum size of DNA packaged by the VLP was determined further. Plasmid DNAs with various sizes were packaged by the VLP in E. coli. Human neuroblastoma cells were then infected with the VLPs containing the various sizes of DNA to allow gene expression. In addition, plasmid DNAs packaged in the VLPs were extracted and retransformed back into E. coli under selection to determine the size of the DNA packaged. The results showed that the JC VLP was able to package plasmid DNA in E. coli up to at least 9.4 kbp in size and this size of DNA could be delivered successfully into human neuroblastoma cells for gene expression. The JC VLP is able to package exogenous DNA up to at least 9.4 kbp in size for gene transduction. These findings will help with the development of gene delivery systems using the JC VLP as the gene delivery vector.

摘要

先前已经证明,JC 病毒样颗粒(VLP)能够在大肠杆菌中包装 DNA,并将 DNA 递送至人类结肠癌细胞中进行基因表达。在本研究中,进一步确定了 VLP 包装的 DNA 的最大尺寸。将具有不同大小的质粒 DNA 包装到大肠杆菌中的 VLP 中。然后,用含有不同大小 DNA 的 VLPs 感染人类神经母细胞瘤细胞,以允许基因表达。此外,从 VLPs 中提取包装的质粒 DNA,并在选择下重新转化回大肠杆菌,以确定包装的 DNA 的大小。结果表明,JC VLP 能够在大肠杆菌中包装至少 9.4 kbp 大小的质粒 DNA,并且该大小的 DNA 可以成功递送至人类神经母细胞瘤细胞中进行基因表达。JC VLP 能够包装外源 DNA ,大小至少为 9.4 kbp ,用于基因转导。这些发现将有助于使用 JC VLP 作为基因传递载体开发基因传递系统。

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