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Matrin 3 Augments the Transcriptional Activity of an SV40 Promoter-Mediated Luciferase Gene with a Highly Repetitive DNA Component

机译:Matrin 3增强了具有高度重复性DNA成分的SV40启动子介导的萤光素酶基因的转录活性。

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摘要

We examined the transcriptional augmentation of matrin 3, a nuclear matrix protein, of the SV40 promotermediated luciferase gene (pGL3) following transient transfection of recombinant plasmids into cells. It has been reported that the interaction of the Xmn I fragment, a highly repetitive DNA component as one of a typical matrix- or scaffold-attachment regions (MAR/SAR) tethered upstream from the SV40 promoter (pGL3-Xmn I) with matrin 3 appeared to be required for augmentation of luciferase gene transcription. In this study, we investigated the levels of induction in cells overexpressing the wild type and several deletion mutants of matrin 3. It appeared that pGL3-Xmn I augmented luciferase production to 4-times the control level in Ac2F cells, but 23-fold in cells overexpressing matrin 3. Electrophoretic mobility shift assay showed that the Xmn I fragment augmented luciferase gene transcription through interaction with matrin 3. Furthermore, our findings suggest that all of the functional domains tested in matrin 3 were necessary for transcriptional augmentation. We aim not only to describe the transcriptional augmentation of matrin 3 with MAR/SAR, but also to strengthen interest in their use to mediate the expression of therapeutic transgenes.
机译:我们检查了重组质粒瞬时转染入细胞后,SV40启动子介导的荧光素酶基因(pGL3)的核基质蛋白matrin 3的转录增强。据报道,Xmn I片段是高度重复的DNA成分,是典型的基质或支架附着区域(MAR / SAR)之一,与SV40启动子(pGL3-Xmn I)上游连接,与基质胶3相互作用。似乎是萤光素酶基因转录增强所必需的。在这项研究中,我们调查了过表达野生型和Matrin 3几个缺失突变体的细胞中的诱导水平。看来pGL3-Xmn I将萤光素酶的产量提高到了Ac2F细胞中对照水平的4倍,而在Ac2F细胞中是23倍。过度表达Matrin 3的细胞。电泳迁移率迁移分析表明Xmn I片段通过与Matrin 3的相互作用增强了荧光素酶基因的转录。此外,我们的发现表明,在Matrin 3中测试的所有功能域对于转录增强都是必需的。我们不仅旨在描述Matrin 3与MAR / SAR的转录增强作用,而且还旨在增强人们对它们介导治疗性转基因表达的兴趣。

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