在体外基因分析Tetrahymena自我拼接内突的基因分析
R Green1, A D Ellington, J W Szostak
1Department of Molecular Biology, Massachusetts General Hospital, Boston 02114.
Nature
|September 27, 1990
概括
研究人员开发了一种新的体外选择和放大系统来分析核酸特性. 这种方法有效地从大型遗传图书馆中分离出活跃的 ribozymes,包括野生类型的变体.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 核酸序列放大使得体外遗传分析成为可能.
- 标准的in vivo方法在选择性条件和变种处理方面存在局限性.
研究的目的:
- 开发一个体外选择和放大系统.
- 使用新系统分析自剪接的Tetrahymena ribozyme.
主要方法:
- 开发了一种新的体外选择和放大系统.
- 应用该系统来研究四胺 ribozyme.
主要成果:
- 在体外系统允许更广泛的选择性条件.
- 它可以处理明显更多的变体,相比于体内的方法.
- 在三个周期中,成功地从超过25万种变体中分离出了利博酶的野生型和活性结构变体.
结论:
- 开发的体外系统对于分析核酸特性非常有效.
- 它能够快速分离功能性 ribozymes 和它们的变体.
- 这种方法比传统的体内基因分析方法具有优势.
相关概念视频
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.


