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Scaffold attachment regions increase reporter gene expression in stably transformed plant cells.

机译:支架附着区增加了稳定转化植物细胞中报告基因的表达。

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摘要

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.
机译:酵母ARS-1元件包含一个支架附着区(SAR),我们之前已经证明它可以在体外与植物核支架结合。为了测试对表达的影响,将嵌合β-葡萄糖醛酸糖苷酶(GUS)基因与该元件侧接的构建体通过微粒轰击传递到烟草悬浮细胞中。在稳定转化的细胞系中,含有两个侧翼SAR的构建体的GUS活性平均比没有SAR的对照构建体高12倍(以基因拷贝为基础,为24倍)。表达水平与基因拷贝数不成正比,如果该元件只是减少位置效应变异所预期的那样。相反,该元素似乎减少了对某些含有多个基因拷贝的转化子中表达的抑制作用。对表达的影响似乎需要染色体整合,因为在瞬时分析中,SAR构建体的活性仅是对照的两倍。

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