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两栖动物卵母细胞中所有主要种类的组蛋白信使核糖核酸的一部分是聚腺苷酸化的。

A portion of all major classes of histone messenger RNA in amphibian oocytes is polyadenylated.

作者信息

Ruderman J V, Pardue M L

出版信息

J Biol Chem. 1978 Mar 25;253(6):2018-25.

PMID:564902
Abstract

Ovaries of the amphibian Triturus viridescens contain a considerable amount of 7 to 12 S RNA which fractionates with poly(A)+ RNA on oligo(dT)-cellulose column chromatography and directs the synthesis of all five classes of histones in a wheat germ cell-free system. The polyadenylate tracts associated with this 7 to 12 S poly (A)+ RNA are heterogeneous in length, ranging from approximately 60 to 120 nucleotides. Partially purified subfractions of histone mRNA templates were isolated from this RNA by preparative polyacrylamide gel electrophoresis. The correlation between the poly(A) content, the template activity in vitro, and the cell-free products encoded by these RNA subfractions suggests that Triturus ovary RNA contains poly(A)+ histone mRNA. The 7 to 12 S poly(A)+ RNA, but not 7 to 12 S poly(A)- RNA, is an effective template for the avian myeloblastosis virus reverse transcriptase-directed synthesis of complementary DNA (cDNA) in the presence of oligo(dT) primer. In the absence of primer, virtually no cDNA is synthesized. When cDNA complementary to 7 to 12 S poly(A)+ RNA is hybridized in situ to Drosophila melanogaster polytene chromosome preparations. cDNA hybrids are found primarily in the region 39D-E, the locus of the histone genes in Drosophila. This cDNA also hybridizes to the sea urchin histone gene sequences contained in the chimeric bacterial plasmids pSp2 and pSp17. These results provide strong evidence for the reality of polyadenylated histone mRNA in Triturus ovary.

摘要

两栖动物绿螈的卵巢含有大量7至12S的RNA,该RNA在寡聚(dT)-纤维素柱层析上与聚(A)+RNA一起分级分离,并在小麦胚无细胞体系中指导所有五类组蛋白的合成。与这种7至12S聚(A)+RNA相关的聚腺苷酸序列长度不均一,范围约为60至120个核苷酸。通过制备性聚丙烯酰胺凝胶电泳从该RNA中分离出组蛋白mRNA模板的部分纯化亚组分。这些RNA亚组分的聚(A)含量、体外模板活性与无细胞产物之间的相关性表明,绿螈卵巢RNA含有聚(A)+组蛋白mRNA。在寡聚(dT)引物存在的情况下,7至12S聚(A)+RNA而非7至12S聚(A)-RNA是禽成髓细胞瘤病毒逆转录酶指导合成互补DNA(cDNA)的有效模板。在没有引物的情况下,几乎不合成cDNA。当与7至12S聚(A)+RNA互补的cDNA原位杂交到黑腹果蝇多线染色体标本上时,cDNA杂交体主要出现在39D-E区域,即果蝇组蛋白基因的位点。该cDNA也与嵌合细菌质粒pSp2和pSp17中包含的海胆组蛋白基因序列杂交。这些结果为绿螈卵巢中聚腺苷酸化组蛋白mRNA的存在提供了有力证据。

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