Investigation on the Interaction between the Molecules Constituting Viruses and Application to Gene Therapy

基因技术的发展和病毒学的基础研究使对人类有害的病毒变成有用的成分成为可能。重组病毒载体在维持哺乳动物细胞体外长期生长的基因转移方法中具有重要的应用价值。采用sv40 -腺病毒重组载体转染人骨髓基质细胞。嵌合病毒载体比DNA转染方法更有效。AAV-2由三种衣壳蛋白组成，比例为VP1: VP2: VP3= 10.0: 1:1。支持AAV-2将基因组封装到预形成的前衣壳中。自组装衣壳的表征及其组装机制对体外系统病毒衣壳的研究具有重要意义。用含有VP2编码区的重组杆状病毒感染昆虫细胞纯化了VLP。从而阐明了参与VLP自组装形成和VLP组分部分解离和重新组装的各种因素。因此，本研究建立了利用新基因载体将重组病毒衣壳蛋白应用于基因治疗的基本研究体系。

It became possible to make viruses which have been hazardous for the human into useful components by the development of gene-technology and basic studies on virology. Recombinant virus vector has been found much worthy in gene transfer method for maintaining the long term growth of mammalian cells in vitro. A SV40-adenovirus recombinant vector was used immortalize the human bone marrow stromal cells. The chimeric virus vector was found more efficient than DNA transfection method. AAV-2, consists of three capsid proteins at a ratio of VP1 : VP2 : VP3=l0 : 1 : 1. AAV-2 is supported to encapsidate the genome into a pre-formed pro-capsid. Characterization of self assembled capsid as well the mechanism of the assembly of the capsid deserve much importance in studying viral capsid for in vitro system. VLP has been purified from insect cells by infecting with recombinant bacurovirus, containing VP2 coding region alone or co-infection. Thus, various factors participating in VLP formation by self assembly and in partial dissociation and re-assembly of VLP components were elucidated. Accordingly, basic study systems of the application of the recombinant virus capsid protein to gene therapy by using vector of novel genes has been established in this study.