Kamiya Hiroyuki, Fukunaga Satoki, Ohyama Takashi, Harashima Hideyoshi
Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Arch Biochem Biophys. 2007 May 1;461(1):7-12. doi: 10.1016/j.abb.2007.02.012. Epub 2007 Mar 8.
The intranuclear disposition of a plasmid is extremely important for transgene expression. The effects of a left-handedly curved sequence with high histone affinity on plasmid expression were examined in vivo. A naked luciferase-plasmid was delivered into mouse liver by a hydrodynamics-based injection, and the luciferase activities were quantitated at various time points. The location of the left-handedly curved sequence determined the transgene expression, without affecting the amount of intranuclear exogenous DNA. The plasmid containing the curved sequence at the location that results in the exposure of the TATA box out of the nucleosome core showed the highest expression. These results suggest that sequences with high histone affinity could control transgene expression from plasmids in vivo.
质粒在细胞核内的分布对于转基因表达极为重要。在体内研究了具有高组蛋白亲和力的左手螺旋弯曲序列对质粒表达的影响。通过基于流体动力学的注射将裸荧光素酶质粒导入小鼠肝脏,并在不同时间点对荧光素酶活性进行定量。左手螺旋弯曲序列的位置决定了转基因表达,而不影响细胞核内外源DNA的量。在导致TATA框暴露于核小体核心之外的位置含有弯曲序列的质粒表现出最高的表达。这些结果表明,具有高组蛋白亲和力的序列可以在体内控制质粒的转基因表达。