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通过DNA过量杂交分析兔脑多核糖体多聚(A+)mRNA

Analysis of rabbit brain polysomal poly (A+) mRNA by DNA excess hybridization.

作者信息

Heikkila J J, Brown I R

出版信息

Biochim Biophys Acta. 1977 Jan 3;474(1):141-53. doi: 10.1016/0005-2787(77)90221-0.

Abstract

DNA sequence representation in rabbit brain mRNA was examined by DNA excess hybridization. purified polysomal poly (A+) mRNA was labeled in vitro with [3H] dimethyl sulfate and reacted at DNA: RNA ratios of 5000 : 1 and 30 000 : 1. Poly(A+) mRNA hybridizied mainly to nonrepeated DNA with a smaller kinetic component which was complementary to repeated DNA sequences. The latter component was not due to nuclear or ribosomal RNA contamination. Poly(A-) RNA purified from brain polysomes hybridized to excess DNA as a single repeated component. Thermal denaturation profiles of the RNA - DNA hybrids indicated a high degree of fidelity in base pairing.

摘要

通过DNA过量杂交检测兔脑mRNA中的DNA序列表现。纯化的多核糖体多聚(A+)mRNA在体外用[3H]硫酸二甲酯标记,并以5000:1和30000:1的DNA:RNA比例反应。多聚(A+)mRNA主要与非重复DNA杂交,有一个较小的动力学成分与重复DNA序列互补。后一成分并非由于核RNA或核糖体RNA污染。从脑多核糖体中纯化的多聚(A-)RNA作为单一重复成分与过量DNA杂交。RNA-DNA杂交体的热变性图谱表明碱基配对具有高度保真度。

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