Allen G C, Hall G E, Childs L C, Weissinger A K, Spiker S, Thompson W F
Department of Botany, North Carolina State University, Raleigh 27695.
Plant Cell. 1993 Jun;5(6):603-13. doi: 10.1105/tpc.5.6.603.
The yeast ARS-1 element contains a scaffold attachment region (SAR) that we have previously shown can bind to plant nuclear scaffolds in vitro. To test effects on expression, constructs in which a chimeric beta-glucuronidase (GUS) gene was flanked by this element were delivered into tobacco suspension cells by microprojectile bombardment. In stably transformed cell lines, GUS activity averaged 12-fold higher (24-fold on a gene copy basis) for a construct containing two flanking SARs than for a control construct lacking SARs. Expression levels were not proportional to gene copy number, as would have been predicted if the element simply reduced position effect variation. Instead, the element appeared to reduce an inhibitory effect on expression in certain transformants containing multiple gene copies. The effect on expression appears to require chromosomal integration, because SAR constructs were only twofold more active than the controls in transient assays.
酵母ARS-1元件包含一个支架附着区域(SAR),我们之前已证明该区域在体外可与植物核支架结合。为了测试其对表达的影响,通过微粒轰击将嵌合β-葡萄糖醛酸酶(GUS)基因两侧带有该元件的构建体导入烟草悬浮细胞。在稳定转化的细胞系中,对于含有两个侧翼SAR的构建体,GUS活性平均比缺乏SAR的对照构建体高12倍(以基因拷贝数计高24倍)。表达水平与基因拷贝数不成比例,而如果该元件只是简单地减少位置效应变异,情况本应如此。相反,该元件似乎减少了对某些含有多个基因拷贝的转化体中表达的抑制作用。对表达的影响似乎需要染色体整合,因为在瞬时测定中,SAR构建体的活性仅比对照高两倍。