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The use of R-looping for structural gene identification and mRNA purification

机译：使用R循环进行结构基因鉴定和mRNA纯化

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摘要

A method is presented for the purification of mRNAs and the identification of structural gene sequences in recombinant DNA molecules. RNA is hybridized to double-stranded linear DNA such that R-loops are formed between most DNAs and their complementary RNA sequences. These R-loops are purified from unhybridized RNAs by gel filtration chromatography in the presence of a high concentration of salt. The complementary RNAs are released from the R-loops by heating, and are assayed by gel electrophoresis or cell free translation to determine their purity and to identify the proteins for which they code. We have demonstrated that recombinant DNAs containing sequences for abundant or moderately abundant mRNAs of Saccharomyces cerevisiae can be identified by this means.
机译：提出了用于纯化MRNA的方法和重组DNA分子中结构基因序列的鉴定。 RNA与双链线性DNA杂交，使得在大多数DNA和它们的互补RNA序列之间形成R环。在高浓度的盐存在下，通过凝胶过滤色谱法从未杂交的RNA纯化这些R环。互补的RNA通过加热从R型环释放，并通过凝胶电泳或无细胞的平移测定以确定它们的纯度并鉴定它们代码的蛋白质。我们已经证明，通过这种方法可以鉴定含有丰富或中等丰富的酿酒酵母MRNA的序列的重组DNA。

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