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通过瞬时表达分析果蝇核糖体DNA启动子

Analysis of the Drosophila rDNA promoter by transient expression.

作者信息

Hayward D C, Glover D M

机构信息

Department of Biochemistry, Imperial College of Science and Technology, London, UK.

出版信息

Nucleic Acids Res. 1988 May 25;16(10):4253-68. doi: 10.1093/nar/16.10.4253.

Abstract

We have examined the expression of the bacterial gene chloramphenicol acetyl transferase (CAT) under the control of the Drosophila rDNA promoter following transfection into Drosophila tissue culture cells. Constructs having an entire NTS, corresponding to approximately 3640 base pairs of upstream rDNA sequence, or constructs with 306 base pairs of upstream sequence respectively, are transcribed at 5 fold or 2 fold higher levels than a construct with 43 base pairs of upstream DNA. In co-transfection experiments, the construct with the entire NTS competes for transcription 20 fold more effectively than the construct with 306 base pairs of upstream sequence. Constructs having either 72 base pairs or 60 base pairs of upstream rDNA sequences, on the other hand, are transcribed very much less efficiently than constructs with either 306 bp or with only 43 bp of upstream DNA. These sequences, which reduce levels of rDNA transcription in the absence of additional upstream DNA, lie in a region in which the rDNA promoter differs from its duplications within the NTS.

摘要

我们检测了转染到果蝇组织培养细胞后,在果蝇核糖体DNA(rDNA)启动子控制下细菌氯霉素乙酰转移酶(CAT)基因的表达情况。含有完整非转录间隔区(NTS)(对应约3640个碱基对的上游rDNA序列)的构建体,或分别含有306个碱基对上游序列的构建体,其转录水平分别比含有43个碱基对上游DNA的构建体高5倍或2倍。在共转染实验中,含有完整NTS的构建体比含有306个碱基对上游序列的构建体在转录竞争上有效20倍。另一方面,含有72个碱基对或60个碱基对上游rDNA序列的构建体,其转录效率比含有306个碱基对或仅43个碱基对上游DNA的构建体低得多。这些在没有额外上游DNA时会降低rDNA转录水平的序列,位于rDNA启动子与其在NTS内的重复序列不同的区域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/77dd/336628/9f5e059b89c5/nar00153-0079-a.jpg

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