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首页> 外文期刊>Molecular reproduction and development >Evaluation of heterologous insulator function with regard to chromosomalposition effect in the mouse blastocyst and fetus
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Evaluation of heterologous insulator function with regard to chromosomalposition effect in the mouse blastocyst and fetus

机译：评估异源绝缘子功能在小鼠胚泡和胎儿中的染色体错配作用

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Insulators are located at the boundaries of differentially regulated genes and delimit their interactions by establishing independent chromatin structures. Recently, an insulator sequence has been found in the 5'-flanking region of arylsulfatase (ARS) gene from sea urchin. To investigate functional conservation of this ARS insulator in mice, we performed blastocyst assays to evaluate the effect of this insulator on the chromosomal position effect, quantitatively. We constructed transgenes that have a luciferase gene under the control of the CMV-IE enhancer and the human elongation factor 1 alpha promoter in the presence or absence of the ARS insulator in both flanking regions, These transgenes were microinjected into 1-cell mouse embryos and luciferase activity was measured at the blastocyst stage. We found that the presence of ARS insulator sequence doubled the number of luciferase-expressing blastocysts, and that the proportion of the blastocysts with high-level expression (greater than or equal to 1 x 10(4) relative light units (RLU)) was increased more than tenfold. In the case of transgenic fetuses, however, the presence of ARS insulator did not seem to improve transgene expression. These results suggest that the sea urchin ARS insulator confers position-independent expression driven by the human elongation factor 1 alpha promoter, at least in the blastocyst stage of the mouse. Mel. Reprod. Dev. 57:232-237, 2000.
机译：绝缘子位于差异调节基因的边界，并通过建立独立的染色质结构来限制它们的相互作用。最近，在海胆的芳基硫酸酯酶（ARS）基因的5'侧翼区域发现了一个绝缘子序列。为了研究这种ARS绝缘子在小鼠中的功能保守性，我们进行了胚泡试验以定量评估这种绝缘子对染色体位置效应的影响。我们构建了在两个侧翼区域都存在或不存在ARS绝缘子的情况下，具有在CMV-IE增强子和人类延伸因子1α启动子控制下的荧光素酶基因的转基因。将这些转基因显微注射到1细胞小鼠胚胎和在胚泡期测量荧光素酶活性。我们发现ARS绝缘子序列的存在使表达萤光素酶的胚泡数目增加了一倍，并且具有高水平表达（大于或等于1 x 10（4）相对光单位（RLU））的胚泡的比例为增加了十倍以上。但是，对于转基因胎儿，ARS绝缘子的存在似乎并未改善转基因表达。这些结果表明，至少在小鼠的胚泡期，海胆ARS绝缘子赋予了由人类延伸因子1α启动子驱动的位置无关的表达。梅尔责备。开发人员57：232-237，2000。

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