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小鼠金属硫蛋白-I基因转染到人类或小鼠细胞后,其转录受镉调控。

The mouse metallothionein-I gene is transcriptionally regulated by cadmium following transfection into human or mouse cells.

作者信息

Mayo K E, Warren R, Palmiter R D

出版信息

Cell. 1982 May;29(1):99-108. doi: 10.1016/0092-8674(82)90094-0.

Abstract

Recombinant vectors containing the mouse metallothionein-I gene (MT-I) and the Escherichia coli xanthine-guanine phosphoribosyltransferase gene (gpt) were used to transfect human hgprt- HeLa cells. Transfected MT-I genes are transcriptionally regulated by cadmium but not by glucocorticoids. S1 mapping indicates that the transcripts from transfected MT-I genes begin at the correct transcription initiation site. We also transfected mouse tk- L cells with a vector containing the mouse MT- I gene and the herpes simplex virus-I thymidine kinase gene. MT-I gene transcription is regulated by cadmium but not by glucocorticoids in this homologous system as well. Finally, we fused the MT-I gene promoter/regulatory region to the thymidine kinase structural gene. Thymidine kinase activity is regulated by cadmium when this fusion gene is transfected into mouse tk- L cells. Deletion mapping experiments indicate that the DNA sequences necessary for regulation of the MT-I gene by cadmium lie within 148 bp of its transcription start site.

摘要

含有小鼠金属硫蛋白-I基因(MT-I)和大肠杆菌黄嘌呤-鸟嘌呤磷酸核糖转移酶基因(gpt)的重组载体被用于转染人hprt-的HeLa细胞。转染的MT-I基因受镉转录调控,但不受糖皮质激素调控。S1图谱分析表明,转染的MT-I基因转录本从正确的转录起始位点开始。我们还用含有小鼠MT-I基因和单纯疱疹病毒-I胸苷激酶基因的载体转染了小鼠tk-L细胞。在这个同源系统中,MT-I基因转录同样受镉调控,而不受糖皮质激素调控。最后,我们将MT-I基因启动子/调控区与胸苷激酶结构基因融合。当这个融合基因转染到小鼠tk-L细胞中时,胸苷激酶活性受镉调控。缺失图谱实验表明,镉对MT-I基因调控所必需的DNA序列位于其转录起始位点的148 bp范围内。

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