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Therapy for Viral Disease by Recombinant Virus or by Recombinant DNA

通过重组病毒或重组DNA治疗病毒性疾病

基本信息

批准号：
13660299
负责人：
KAI Kazushige
金额：
$ 2.56万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

The virus vector system that transduces some genes simultaneously seems to be very useful for various gene transducing therapy. In this research, we intended to develop a retrovirus vector that carries two genes in the gag and the env gene regions of Friend murine leukemia virus (F-MuLV). We madevariousvectors that lost gag initiation codons, bare various types of Human Cytomegalovirus (CMV) promoter in stead of the enhancer sequence of LTR of MULV, and/or bare EGFP gene in the gag gene region and H- or F-gene of canine distemper virus (CDV) in the env gene region of MuLV. Results are followings; 1)The lost of initiation codons did not affect the expression of introduced gene in gag gene region, 2)The introduction of CMV promotor was necessary for the expression of introduced genes in the xeno-genic cells than mouse or rat cells although all of introduced segments from CMV promotor showed no difference among them (manuscript in preparation), 3)Both gag gene and env gene regions were responsible for introducing exogenous genes, 4)The H gene of CDV induced cytopathic effect and the F gene of CDV iduced cell-fusion on transfected Vero cells, 5)The co-transfection of both H- or F-bearing vector DNA showed slow progress of cell fusion and the process of cell fusion was possibly monitered by the introduced EGFP gene product (manuscript in preparation).

同时转导多个基因的病毒载体系统在各种基因转导治疗中有着广泛的应用前景。在本研究中，我们打算开发一个逆转录病毒载体，携带两个基因的gag和env基因区的朋友小鼠白血病病毒（F-MuLV）。我们制作了多种载体，这些载体丢失了gag起始密码子，裸了不同类型的人巨细胞病毒（CMV）启动子，而不是MULV的LTR的增强子序列，和/或裸了MuLV的gag基因区的EGFP基因和犬瘟热病毒（CDV）的H-或F-基因。结果如下：1）gag基因区起始密码子的丢失并不影响外源基因的表达; 2）外源基因在异种细胞中的表达需要CMV启动子的导入，而在小鼠和大鼠细胞中的表达则需要CMV启动子的导入，尽管它们之间的CMV启动子的导入片段没有差异3）gag基因和env基因区域都负责外源基因的导入，4）CDV的H基因在转染的Vero细胞上引起细胞病变，CDV的F基因在转染的Vero细胞上引起细胞融合，5）共转染H-或F-携带载体DNA的细胞融合进展缓慢，细胞融合过程可能通过引入的EGFP基因产物进行监控（手稿在制备中）。