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大鼠肝脏不均一核RNA中铜蓝蛋白信使RNA序列的鉴定。

Identification of ceruloplasmin messenger RNA sequences in heterogeneous nuclear RNA from rat liver.

作者信息

Gaitskhoki V S, L'vov V M, Schwartzman A L, Skobeleva N A, Frolova L J, Neifakh S A

出版信息

Mol Biol Rep. 1981 Nov 30;8(1):57-62. doi: 10.1007/BF00798386.

Abstract

The distribution of the sequences of ceruloplasmin mRNA in different fractions of heterogeneous nuclear RNA from rat liver was studied using cDNA transcripts of highly purified mRNA as hybridization probe. The content of ceruloplasmin mRNA sequences in poly(A)-containing and poly(A)-free subfractions of heterogeneous nuclear RNA is respectively 1 and 27 molecules per a hepatocyte. Heterogeneous nuclear RNA carrying the sequences of ceruloplasmin mRNA sedimented in sucrose gradients containing formamide, as a broad zone around the 56S peak. Denaturing electrophoresis followed by the transfer of RNA onto diabenzyloxymethyl paper and hybridization with [32P]-cDNA revealed multiple high molecular weight fractions of ceruloplasmin pre = mRNA (9.0, 6.6, 2.2 and 1.6 megadaltons) in the non-adenylated fraction of nuclear RNA and a single 1.1-1.2 megadalton zone in poly(A)-containing nuclear RNA, the latter being equal in size to the mature ceruloplasmin mRNA from liver polysomes.

摘要

使用高度纯化的mRNA的cDNA转录本作为杂交探针,研究了大鼠肝脏异质核RNA不同组分中铜蓝蛋白mRNA序列的分布。异质核RNA中含poly(A)和不含poly(A)的亚组分中铜蓝蛋白mRNA序列的含量分别为每个肝细胞1个分子和27个分子。携带铜蓝蛋白mRNA序列的异质核RNA在含有甲酰胺的蔗糖梯度中沉降,形成一个围绕56S峰的宽区带。变性电泳后将RNA转移到二苄氧基甲基纸上,并与[32P]-cDNA杂交,结果显示核RNA的非腺苷化组分中有多个高分子量的铜蓝蛋白前体mRNA组分(9.0、6.6、2.2和1.6兆道尔顿),而含poly(A)的核RNA中有一个单一的1.1 - 1.2兆道尔顿区带,后者的大小与来自肝脏多核糖体的成熟铜蓝蛋白mRNA相同。

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