Establishment of gene therapy vector by using DNA replication origin and virus vector

利用DNA复制起点和病毒载体建立基因治疗载体

• 批准号: 07557147
• 负责人: ARIGA Hiroyoshi
• 金额: $ 8.7万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557147/
• 关键词: MYC; adenovirus; vector; heat shock protein; transcription; DNA replication; c-myc; DNA複製開始; 結合タンパク質; 癌遺伝子; DNA結合; 転写因子; 発現ベクター; 細胞周期

Expression vector composed of adenovirus, beta-galactosidase, and DNA replication origin (ori) was constricted.Oris used here were from genes of c-myc, human hsp70 and immunoglobulin heavy chain in addition to SV40 origin as a positive control. These origins were inserted to adenovirus vector containing Cre-lox system, and the recombinant adenovirus were recovered from human 2983 cells transfected with these vectors. Adenovirus containing SV40 origin were first infected to monkey CosI cells, and the circular adenovirus expressing beta-galactosidase was recovered. Adenovirus containing hsp70 gene origin were then infected to human HeLa cells, and the circular form of adenovirus DNA was also recovered. Copy number of hsp70 ori derived plasmid DNA in the cells, however, was lower than that of SV40 origin in 10^<-3> order. Results suggest that this system should be useful for an expression vector for gene therapy.

由腺病毒，β-半乳糖苷酶和DNA复制起源（ORI）组成的表达载体被限制。此处使用的ORIS来自C-Myc，Human HSP70和免疫球蛋白重链的基因，除了SV40外，还来自SV40起源。将这些起源插入含有Cre-Lox系统的腺病毒载体，并从用这些载体转染的人类2983细胞中回收重组腺病毒。首先将含有SV40起源的腺病毒病毒感染到猴子COSI细胞中，并回收表达β-半乳糖苷酶的圆形腺病毒。然后将含有HSP70基因起源的腺病毒感染到人的HeLa细胞中，并回收了腺病毒DNA的圆形形式。然而，细胞中HSP70衍生的质粒DNA的拷贝数低于10^<-3>顺序中的Sv40原点。结果表明，该系统对于基因治疗的表达载体应该有用。

项目成果

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