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首页> 外文期刊>Journal of Virological Methods >Display of Hepatitis B Virus PreS1 peptide on bacteriophage T7 and its potential in gene delivery into HepG2 cells
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Display of Hepatitis B Virus PreS1 peptide on bacteriophage T7 and its potential in gene delivery into HepG2 cells

机译：乙型肝炎病毒PreS1肽在噬菌体T7上的展示及其在基因导入HepG2细胞中的潜力

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Hepatitis B is a major public health problem worldwide which may lead to chronic liver diseases, cirrhosis and hepatocellular carcinoma. An interaction between hepatitis B virus (HBV) envelope protein, particularly the PreS1 region, and a specific cell surface receptor is believed to be the initial step of HBV infection through attachment to hepatocytes. In order to develop a gene delivery system, bacteriophage T7 was modified genetically to display polypeptides of the PreS1 region. A recombinant T7 phage displaying amino acids 60-108 of the PreS1 region (PreS1 sub(6) sub(0) sub(-) sub(1) sub(0) sub(8)) was demonstrated to be most effective in transfecting HepG2 cells in a dose- and time-dependant manner. The phage genome was recovered from the cell lysate and confirmed by PCR whereas the infectious form of the internalized phage was measured by a plaque-forming assay. The internalized phage exhibited the appearance of green fluorescent dots when examined by immunofluorescence microscopy. Surface modification, particularly by displaying the PreS1 sub(6) sub(0) sub(-) sub(1) sub(0) sub(8) enhanced phage uptake, resulting in more efficient in vitro gene transfer. The ability of the recombinant phage to transfect HepG2 cells demonstrates the potential of the phage display system as a gene therapy for liver cancer.
机译：乙型肝炎是世界范围内的主要公共卫生问题，可能导致慢性肝病，肝硬化和肝细胞癌。乙型肝炎病毒（HBV）包膜蛋白（特别是PreS1区）与特定细胞表面受体之间的相互作用被认为是通过附着于肝细胞感染HBV的第一步。为了开发基因递送系统，对噬菌体T7进行了遗传修饰以展示PreS1区域的多肽。展示了显示PreS1区60-108位氨基酸（PreS1 sub（6）sub（0）sub（-）sub（1）sub（0）sub（8）的重组T7噬菌体对HepG2的转染最有效细胞具有剂量和时间依赖性。从细胞裂解物中回收噬菌体基因组并通过PCR确认，而通过噬菌斑形成测定法来测定内化噬菌体的感染形式。当通过免疫荧光显微镜检查时，内化的噬菌体表现出绿色荧光点的外观。表面修饰，特别是通过显示PreS1 sub（6）sub（0）sub（-）sub（1）sub（0）sub（8）可以增强噬菌体的摄取，从而提高体外基因转移的效率。重组噬菌体转染HepG2细胞的能力证明了该噬菌体展示系统作为肝癌基因治疗的潜力。

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