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腺相关病毒(Adeno-associatedvirus,AAV)是一类线状单股DNA(SS-DNA)的微小病毒。依据Cavalier-Smith的SS-DNA病毒复制模型原理,将其末端反向重复顺序(ITRs)分离,变为折叠十字架结构,再以其发夹中的3'-OH为引物合成另一股姊妹染色体DNA,然后共价连接到亲代分子上,与辅助病毒Ad2和PAAV/Ad共转染真核生物细胞。Southern印迹分子杂交结果表明:插入这一折叠结构中由SV40早期启动子控制的报告基因-新霉素抗性基因拷贝已整合到细胞基因组染色体上,这种以无细菌DNA分子为支架的折叠结构不仅证明了Cavalier-Smith的理论,而且可望为外源基因导入真核细胞提供新的又一系统。
Adeno-associated virus (AAV) is a type of linear virus (SS-DNA) parvovirus. According to Cavalier-Smith’s principle of SS-DNA virus replication model, ITRs were separated into folded crosses, and the other 3’-OH was used as a primer to synthesize another sister chromosome DNA, and then covalently attached to the parental molecule, co-transfecting eukaryotic cells with the helper viruses Ad2 and PAAV / Ad. The results of Southern blot hybridization showed that the copy of neomycin resistance gene, a reporter gene controlled by the SV40 early promoter inserted into this folded structure, has been integrated into the genome of the cell genome. This folding structure with bacterial DNA as a scaffold It not only proved the theory of Cavalier-Smith, but also expected to provide a new system for the introduction of foreign genes into eukaryotic cells.