中文摘要：

噬菌体整合酶ΦBT1因其具有可介导转基因位点特异性整合的能力而成为了基因治疗中一种有效的工具,它丰富了转基因载体的选择,并使得多位点特异性整合成为可能.为了能够更加安全有效地利用ΦBT1整合酶作为基因治疗载体,有必要了解ΦBT1整合酶与宿主细胞内蛋白质相互作用的情况.酵母配对实验与免疫共沉淀实验揭示了ΦBT1整合酶中4个氨基酸433RFAL436对整合酶与PML-NBs蛋白Daxx的结合起到了关键作用.通过进一步构建并利用ΦBT1整合酶哺乳动物细胞报告系统,证实过表达Daxx会抑制ΦBT1整合酶在293T细胞中的重组效率.以上结果表明,细胞内的蛋白质可以与ΦBT1整合酶发生相互作用并抑制其重组活性,对于改善ΦBT1整合酶介导的转基因操作以及选择ΦBT1整合酶靶细胞方面具有重要的参考意义.

英文摘要：

The bacterial phage ΦBT1 integrase is a promising tool due to its site-specific transgene character.It enriches the site-specific transgenic tools and provides the possibility for multiple site-specific transgenic manipulations.To improve its safety as a vector of gene therapy,it is necessary to investigate the potential interactions between ΦBT1 and proteins in mammalian host cells.Yeast mating and co-immunoprecipitation assay indicated that a tetrapeptide 433RFAL436 in ΦBT1 integrase was responsible for ΦBT1 and Daxx interaction.It was also demonstrated that over-expression of Daxx could reduce ΦBT1 mediated recombination rate in 293T cells by using ΦBT1 report system.It is the first time to identify a cellular protein interacting with ΦBT1 integrase and inhibiting its recombination efficiency.This result might be useful for improving the ΦBT1 integrase mediated transgene methods and directing the selection of target cells for ΦBT1 integrase.