New rapid gel sequencing method for RNA
New rapid gel sequencing method for RNA
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新型 RNA 快速凝胶测序方法
作者：
A. Simoncsits;G. Brownlee;R. Brown;J. Rubin;H. Guilley
由于快速“读出”凝胶方法的发展，核酸测序方法在过去几年中有了显着改善。第一种被开发出来的方法是dna的“正负”复制方法，现在也存在一种类似的RNA复制方法。一种不同的、更直接的DNA测序方法已经开发出来，但还没有对RNA进行直接测序的方法。我们已经开发了这样一种方法，在这里我们描述了它在建立嗜热脂肪芽孢杆菌酪氨酸tRNA序列中的应用。这个序列特别有趣，因为B. stearthermoophilus中的酪氨酸tRNA及其特异性合成酶为研究tRNA和合成酶的特异性相互作用提供了一个有吸引力的实验系统，这是由于x射线晶体学研究蛋白质三维结构的进展而成为可能的。
METHODS for sequencing nucleic acids have improved significantly in the last few years due to the development of rapid ‘read-off’ gel methods. The first method to be developed was the ‘plus and minus’ copying method for DNA1 and an analogous copying method for RNA now exists2. A different and more direct method for sequencing DNA has been developed3, but no direct method is available for RNA. We have developed such a method and here we describe its use in establishing the sequence of tyrosine tRNA of Bacillus stearothermophilus. This sequence is especially interesting as the tyrosine tRNA and its specific synthetase in B. stearothermophilus offer an attractive experimental system for studying the specific interaction of tRNA and synthetases, which is now possible because of progress in studying the three-dimensional structure of the protein by X-ray crystallography4.