相关实验视频
Updated: May 28, 2026

11:48
Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
Published on: October 9, 2014
纯化前体的体外蛋白质拼接和分支中间体的识别
M Q Xu1, M W Southworth, F B Mersha
1New England Biolabs, Beverly, Massachusetts 01915.
Cell
|December 31, 1993
概括
这项研究提供了蛋白质拼接的直接证据,这是一个删除蛋白质部分并连接剩余部分的过程. 在体外实验表明,这种拼接机制取决于温度和pH值.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 蛋白质化学 蛋白质化学
背景情况:
- 蛋白质拼接是一种关键的翻译后修改.
- 它涉及切除内部聚和绑定侧面序列的切除.
- 这一过程的结果是从单个前体中形成两个不同的蛋白质.
研究的目的:
- 为蛋白质拼接提供直接证据.
- 为了研究一种含有Pyrococcus DNA聚合酶拼接元件的蛋白质前体的体外拼接.
- 描述影响蛋白质拼接的条件.
主要方法:
- 从Pyrococcus DNA聚合酶中克隆蛋白质拼接元件到异物基因中.
- 净化已积累的前体蛋白质.
- 在体外进行拼接试验.
- 使用N-终端测序分析拼接中间体.
- 研究温度和pH对拼接的影响.
主要成果:
- 在实验室中证明了蛋白质拼接的直接证据.
- 发现拼接活动取决于温度和pH值.
- 一种缓慢迁移的物种,通过N-终端测序被确定为一个分支分子,表现出拼接中间体的特征.
- 前体和中间体物种在pH值变化时可以相互转换.
结论:
- 蛋白质拼接可以在体外证明和研究.
- 温度和pH值是调节蛋白质拼接效率的关键因素.
- 一种分支的中间体参与了蛋白质拼接机制.
- 这些发现有助于理解蛋白质加工的分子机制.
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...
pre-mRNA Processing
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
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...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Pre-mRNA Processing: 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...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...

