开发一种基于基的净化系统,利用基结合域和烟草蚀刻病毒蛋白酶裂变,以有效地恢复重组蛋白质
1Department of Biotechnology and Food Technology, Southern Taiwan University of Science and Technology, Tainan 710, Taiwan.
Journal of agricultural and food chemistry
|September 28, 2024
概括
这项研究介绍了一种基于基的新型蛋白质净化系统,在超折叠绿色光蛋白 (sfGFP) 和三合成酶 (TaTS) 等目标蛋白质中达到90%以上的纯度. 该方法有效地去除不需要的蛋白质,用于工业应用.
科学领域:
- 生物化学 生物化学
- 蛋白质净化 蛋白质净化
- 生物技术是生物技术.
背景情况:
- 素结合域 (ChBD) 对于蛋白质固定是有效的.
- 蛋白酶分裂是释放纯化蛋白质的常见方法.
- 高效的净化系统对于重组蛋白质生产至关重要.
研究的目的:
- 开发一种有效的基基净化系统,用于准蛋白质.
- 利用一种新的策略,涉及特定的蛋白酶裂变和二次素吸附.
- 为了实现重组蛋白质的高纯度,特别是sfGFP和TaTS.
主要方法:
- 蛋白质 (sfGFP,TaTS) 与素结合域 (ChBD) 和烟草蚀刻病毒蛋白酶 (TEVp) 分裂部位融合.
- 目标蛋白质被固定在基珠上.
- 使用突变TEVp (H-TEVS219V-ChBD) 的目标蛋白的裂变.
- 二次净化步骤使用新鲜的丁珠来去除分裂的蛋白酶.
主要成果:
- 这种基于基的系统成功地固定和净化了sfGFP和TaTS.
- 对于两个标蛋白质,实现了超过90%的电泳级纯度.
- 该系统证明了能有效地去除内源性和分裂的蛋白质酶污染物.
结论:
- 开发的基于基的净化系统是高效和特定的.
- 这种方法为工业规模的蛋白质净化提供了一个有前途的方法.
- 该系统可以达到高纯度水平,适合各种应用.
相关概念视频
Conservative Site-specific Recombination and Phase Variation
5.7K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
5.7K
Tagging and Fusion Proteins
6.4K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
6.4K


