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卵清蛋白基因中结构序列和间隔序列的转录以及潜在卵清蛋白mRNA前体的鉴定。

Transcription of structural and intervening sequences in the ovalbumin gene and identification of potential ovalbumin mRNA precursors.

作者信息

Roop D R, Nordstrom J L, Tsai S Y, Tsai M J, O'Malley B W

出版信息

Cell. 1978 Oct;15(2):671-85. doi: 10.1016/0092-8674(78)90035-1.

Abstract

Structural sequences that are extensively separated by nonstructural intervening sequences in the natural ovalbumin gene are coordinately expressed in target and nontarget tissue. The intervening sequences, which consist of unique sequences in the chick genome, are transcribed in their entirety. The amount of nuclear RNA corresponding to these sequences, however, is approximately 10 times less than that observed for structural sequences. The accumulation of RNA corresponding to structural and intervening sequences during acute estrogen stimulation suggests either that there are different rates of transcription for these regions of the ovalbumin gene or that RNA sequences corresponding to the intervening sequences are preferentially processed and degraded. Comparison of the in vitro expression of portions of the ovalbumin gene in nuclei isolated from chronically stimulated oviducts indicates that both structural and intervening sequences are preferentially transcribed in vitro at rates approximately 500 times greater than expected for random transcription of the haploid chick genome. In addition, electrophoresis of oviduct nuclear RNA on agarose gels containing methylmercury hydroxide reveals multiple species of RNA that are from 1.3 to over 4 times larger than ovalbumin mRNA and hybridize to both structural and intervening sequences of the ovalbumin gene. These results are consistent with transcription of the entire ovalbumin gene into a large precursor molecule followed by excision of the intervening sequences and appropriate ligation of the structural sequences to form the mature mRNA.

摘要

在天然卵清蛋白基因中被非结构间隔序列广泛分隔的结构序列,在靶组织和非靶组织中协同表达。这些间隔序列由鸡基因组中的独特序列组成,会被完整转录。然而,与这些序列对应的核RNA量大约比结构序列观察到的量少10倍。急性雌激素刺激期间与结构和间隔序列对应的RNA积累表明,要么卵清蛋白基因这些区域的转录速率不同,要么与间隔序列对应的RNA序列被优先加工和降解。对从长期受刺激的输卵管分离的细胞核中卵清蛋白基因部分的体外表达进行比较表明,结构和间隔序列在体外均被优先转录,其速率比单倍体鸡基因组随机转录预期的速率大约高500倍。此外,在含有氢氧化甲基汞的琼脂糖凝胶上对输卵管核RNA进行电泳,发现多种RNA,其大小比卵清蛋白mRNA大1.3至4倍以上,并与卵清蛋白基因的结构和间隔序列杂交。这些结果与整个卵清蛋白基因转录成一个大的前体分子,随后切除间隔序列并将结构序列进行适当连接以形成成熟mRNA一致。

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