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与小鼠L细胞中重复DNA互补的多聚腺苷酸化RNA。

Polyadenylated RNA complementary to repetitive DNA in mouse L-cells.

作者信息

Ryffel G U, McCarthy B J

出版信息

Biochemistry. 1975 Apr 8;14(7):1385-9. doi: 10.1021/bi00678a007.

Abstract

Complementary DNA, synthesized with L-cell polyadenylated RNA as template, renatured with total L-cell DNA to about 70%. About 30% complementary to unique sequence DNA and another 10 and 30% corresponded to sequences about 20- and 500-fold repetitive. Complementary DNA was fractionated after partial hybridization with total polyadenylated RNA to obtain preparations enriched or impoverished in complements of the most frequent polyadenylated RNA. Renaturation of these complementary DNA fractions with L-cell DNA revealed that most frequent RNAs are transcribed from repetitive DNA sequences, Complementary DNA, density labeled with bromodeoxyuridine, was fractionated by renaturation with L-cell DNA to yield fractions enriched in repetitive and unique sequence DNA. The denisty labeled complementary DNA was purified by equilibrium centrifiguation in an alkaline Cs2SO4 gradient. The complementary DNA representing mainly repetitive DNA sequences hybridized preferentially to frequent polyadenylated RNA.

摘要

以L细胞多聚腺苷酸化RNA为模板合成的互补DNA,与总L细胞DNA复性至约70%。约30%与单一序列DNA互补,另外10%和30%分别对应约20倍和500倍重复序列。与总多聚腺苷酸化RNA进行部分杂交后,对互补DNA进行分级分离,以获得富含或贫化最常见多聚腺苷酸化RNA互补物的制剂。这些互补DNA级分与L细胞DNA复性表明,最常见的RNA是从重复DNA序列转录而来的。用溴脱氧尿苷进行密度标记的互补DNA,通过与L细胞DNA复性进行分级分离,以产生富含重复序列和单一序列DNA的级分。通过在碱性Cs2SO4梯度中进行平衡离心,对密度标记的互补DNA进行纯化。主要代表重复DNA序列的互补DNA优先与常见的多聚腺苷酸化RNA杂交。

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