Namciu S J, Blochlinger K B, Fournier R E
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109-1024, USA.
Mol Cell Biol. 1998 Apr;18(4):2382-91. doi: 10.1128/MCB.18.4.2382.
Germ line transformation of white- Drosophila embryos with P-element vectors containing white expression cassettes results in flies with different eye color phenotypes due to position effects at the sites of transgene insertion. These position effects can be cured by specific DNA elements, such as the Drosophila scs and scs' elements, that have insulator activity in vivo. We have used this system to determine whether human matrix attachment regions (MARs) can function as insulator elements in vivo. Two different human MARs, from the apolipoprotein B and alpha1-antitrypsin loci, insulated white transgene expression from position effects in Drosophila melanogaster. Both elements reduced variability in transgene expression without enhancing levels of white gene expression. In contrast, expression of white transgenes containing human DNA segments without matrix-binding activity was highly variable in Drosophila transformants. These data indicate that human MARs can function as insulator elements in vivo.
用含有白色基因表达盒的P因子载体对白果蝇胚胎进行种系转化,由于转基因插入位点的位置效应,会导致果蝇出现不同的眼色表型。这些位置效应可以被特定的DNA元件消除,比如在体内具有绝缘子活性的果蝇scs和scs'元件。我们利用这个系统来确定人类基质附着区域(MARs)在体内是否能作为绝缘子元件发挥作用。来自载脂蛋白B和α1-抗胰蛋白酶基因座的两种不同的人类MARs,使白色转基因表达免受黑腹果蝇位置效应的影响。这两种元件都降低了转基因表达的变异性,而没有提高白色基因的表达水平。相比之下,含有无基质结合活性的人类DNA片段的白色转基因在果蝇转化体中的表达高度可变。这些数据表明,人类MARs在体内可以作为绝缘子元件发挥作用。