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导入小鼠红白血病细胞中的β-珠蛋白基因的调控表达。

The regulated expression of beta-globin genes introduced into mouse erythroleukemia cells.

作者信息

Chao M V, Mellon P, Charnay P, Maniatis T, Axel R

出版信息

Cell. 1983 Feb;32(2):483-93. doi: 10.1016/0092-8674(83)90468-3.

Abstract

We have introduced a hybrid mouse-human beta-globin gene as well as the intact human beta-globin gene into murine erythroleukemia (MEL) cells and have demonstrated that these genes are appropriately regulated during differentiation of the MEL cell in culture. The addition of chemical inducers to cotransformed cells results in a 5 to 50 fold increase in the level of mRNA transcribed from the exogenous globin gene. S1 nuclease and primer extension analyses demonstrate that these mRNAs initiate and terminate correctly. Nuclear transcription experiments indicate that induction of hybrid mRNA results at least in part from the increase in the rate of globin gene transcription. Furthermore, the induction appears to be specific for globin genes within an erythroid cell. These results permit the study of expression of the globin gene during erythroid differentiation and suggest that the specific induction of the globin gene is an inherent property of DNA sequences within or flanking the beta-globin genes. Moreover, the fact that the human and hybrid globin genes are both inducible in MEL cells suggests that these regulatory sequences are conserved between mouse and human cells.

摘要

我们已将一个杂交的小鼠 - 人β - 珠蛋白基因以及完整的人β - 珠蛋白基因导入鼠红细胞白血病(MEL)细胞,并证明这些基因在培养的MEL细胞分化过程中受到适当调控。向共转化细胞中添加化学诱导剂会使外源珠蛋白基因转录的mRNA水平增加5至50倍。S1核酸酶和引物延伸分析表明这些mRNA起始和终止正确。核转录实验表明,杂交mRNA的诱导至少部分源于珠蛋白基因转录速率的增加。此外,这种诱导似乎对红系细胞内的珠蛋白基因具有特异性。这些结果有助于研究红系分化过程中珠蛋白基因的表达,并表明珠蛋白基因的特异性诱导是β - 珠蛋白基因内部或侧翼DNA序列的固有特性。此外,人和杂交珠蛋白基因在MEL细胞中均可诱导这一事实表明,这些调控序列在小鼠和人类细胞之间是保守的。

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