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注射到非洲爪蟾卵母细胞核中的组蛋白基因的转录保真度。

Transcriptional fidelity of histone genes injected into Xenopus oocyte nuclei.

作者信息

Hentschel C, Probst E, Birnstiel M L

出版信息

Nature. 1980 Nov 6;288(5786):100-2. doi: 10.1038/288100a0.

Abstract

Previous work has indicated that at least some of the genetic information required for the expression of sea urchin histone genes is recognized following injection of the gene repeat (h22) into Xenopus oocyte nuclei. The ability to elicit the expression of cloned genes and their sequence-manipulated counterparts is proving invaluable in analysing the molecular details of gene expression. Direct injection of such genes into Xenopus oocyte nuclei remains one of the simplest methods for obtaining such expression and a remarkable degree of transcriptional fidelity has been demonstrated using this system with RNA polymerase III genes, and to a lesser extent with rDNAs transcribed by RNA polymerase I. In the case of polymerase II genes there is ample evidence for coupled transcription-translation, but the degree of transcriptional fidelity involved may, as has recently been shown for the ovalbumin gene, be minimal. However, clearly if the oocyte is to be used to investigate transcriptional regulation of such genes, transcriptional fidelity defined as the production of correct RNA termii, rather than the production of 'functional mRNAs' (ref. 15), must pertain. Here we demonstrate such fidelity in the expression of all five Psammechinus miliaris histone genes comprising a repeat unit. However, we find large quantitative variations in the levels of synthesis of the individual correct termini and hence of the mRNAs. In addition to the mRNAs, species with no detectable counterparts in the sea urchin are generated off the coding strand, as are heterogeneous noncoding species.

摘要

先前的研究表明,将海胆组蛋白基因重复序列(h22)注入非洲爪蟾卵母细胞核后,至少能识别出部分表达所需的遗传信息。在分析基因表达的分子细节方面,诱导克隆基因及其经序列操作的对应物表达的能力已被证明具有极高的价值。将此类基因直接注入非洲爪蟾卵母细胞核仍然是获得这种表达的最简单方法之一,并且使用该系统对RNA聚合酶III基因进行研究时,已证明具有相当高的转录保真度,对RNA聚合酶I转录的rDNA的研究也在一定程度上证明了这一点。对于聚合酶II基因,有充分的证据表明存在转录与翻译的偶联,但正如最近对卵清蛋白基因的研究所显示的那样,所涉及的转录保真度可能很低。然而,显然,如果要利用卵母细胞来研究此类基因的转录调控,那么定义为产生正确RNA末端而不是“功能性mRNA”(参考文献15)的转录保真度必须存在。在这里,我们证明了包含一个重复单元的所有五个沙钱组蛋白基因在表达时具有这种保真度。然而,我们发现各个正确末端以及由此产生的mRNA的合成水平存在很大的定量差异。除了mRNA之外,在编码链上还产生了在海胆中无法检测到对应物的物种,以及异质的非编码物种。

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