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将克隆的完整基因注射到非洲爪蟾卵母细胞核后观察到的鸡卵清蛋白基因转录单位。

Transcription units of chicken ovalbumin gene observed after injection of cloned complete genes into Xenopus oocyte nuclei.

作者信息

Trendelenburg M F, Mathis D, Oudet P

出版信息

Proc Natl Acad Sci U S A. 1980 Oct;77(10):5984-8. doi: 10.1073/pnas.77.10.5984.

Abstract

The organization of transcription of a well-characterized protein encoding gene was studied by microinjection and electron microscopy. Circular recombinant DNA molecules containing the complete chicken ovalbumin sequences (7.7 kilobases, contained in 11.5 kilobases of chicken DNA) were microinjected into germinal vesicles of living oocytes of the clawed toad Xenopus laevis, and their transcription was studied in nuclear spread preparations. Evaluation of spread chromatin showed a limited number of observed molecules transcribed in "specific" patterns--i.e., circular chromatin molecules containing transcription units approximately 2.3 micrometer long, consisting of regular series of densely packed lateral ribonucleoprotein fibrils gradually increasing in length. The appearance of these fibril gradients was similar to that of actively transcribed endogenous protein encoding genes contained in lampbrush-chromosome loops of the same nuclei and to the putative Bombyx silk fibroin transcription units. In addition, less-regular arrays of transcript fibrils were seen in some circles, including fully fibril-covered molecules, indicative of the occurrence of irregular transcriptional events. The results of this heterologous transcription experiment indicate that the transcriptional machinery of the amphibian oocyte nucleus is capable of transcribing protein encoding genes from an avian species in typical regular arrays of transcription units.

摘要

通过显微注射和电子显微镜技术,对一个特征明确的蛋白质编码基因的转录组织进行了研究。将含有完整鸡卵清蛋白序列(7.7千碱基,包含在11.5千碱基的鸡DNA中)的环状重组DNA分子显微注射到非洲爪蟾(Xenopus laevis)活卵母细胞的生发泡中,并在核铺展制剂中研究其转录情况。对铺展染色质的评估显示,观察到的分子中只有少数以“特定”模式转录,即含有约2.3微米长转录单位的环状染色质分子,由一系列规则排列的紧密堆积的侧向核糖核蛋白纤维组成,其长度逐渐增加。这些纤维梯度的外观与同一细胞核灯刷染色体环中所含的活跃转录的内源性蛋白质编码基因以及家蚕丝心蛋白推定转录单位的外观相似。此外,在一些环中还观察到不太规则的转录纤维阵列,包括完全被纤维覆盖的分子,这表明存在不规则的转录事件。这个异源转录实验的结果表明,两栖类卵母细胞核的转录机制能够以典型的规则转录单位阵列转录鸟类物种的蛋白质编码基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c62b/350197/f1ea5dd44994/pnas00497-0446-a.jpg

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