Animal Genome Research Unit, Division of Animal Science, National Institute of Agrobiological Sciences, 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan.
N Biotechnol. 2011 Jul;28(4):312-9. doi: 10.1016/j.nbt.2010.11.001. Epub 2010 Nov 11.
phiC31 integrase-based gene delivery has been developed. However, the expression of integrated transgenes is often suppressed by a negative position effect. To improve this system, we constructed a new phiC31 integrase-based expression vector that contains attB, an expression unit placed in reverse orientation with two sea urchin-derived Ars-insulators to avoid position effects. In vitro and in vivo transfection experiments revealed that this new system produces higher levels of transgene expression as well as continued gene expression. Thus, the present gene delivery system will facilitate reverse genetics-based molecular biological studies.
phiC31 整合酶基因传递系统已经建立。然而，整合转基因的表达常常受到负位置效应的抑制。为了改进该系统，我们构建了一种新型 phiC31 整合酶表达载体，该载体包含 attB，其表达单元以反向方式与两个海胆衍生的 Ars-insulator 排列，以避免位置效应。体外和体内转染实验表明，该新系统可产生更高水平的转基因表达以及持续的基因表达。因此，该基因传递系统将有助于基于反向遗传学的分子生物学研究。
