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Updated: Jan 29, 2026

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High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
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洞察CusF作为重组蛋白表达的可溶性标签的性能
Igor P Oscorbin1,2, Maria A Smertina1, Maria S Kunova1,2
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS), 8, Lavrentiev Avenue, Novosibirsk 630090, Russia.
International journal of molecular sciences
|January 28, 2026
概括
CusF 蛋白标签并没有改善两种测试的重组蛋白质的可溶性或铜 afinity 净化. 需要进一步的研究才能找到有效的蛋白质可溶性标签.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 蛋白质工程是指蛋白质工程.
背景情况:
- CusF蛋白是一种金属结合的周等离子体蛋白.
- 它已被提议作为一种双功能标签,用于增强重组蛋白溶解性,并使铜亲和染色学成为可能.
- 证据证明CusF作为标签的表现有限.
研究的目的:
- 为了评估CusF作为两个异质蛋白质的溶解性标签:Efa PAP和mCherry.
- 评估CusF提高蛋白质可溶性和通过IMAC促进净化的能力.
- 扩大关于可溶性标签性能的经验知识.
主要方法:
- 合CusF到Efa PAP和mCherry. 这是一个很好的方法.
- 在大肠杆菌菌株中复合蛋白的表达 (BL21 (DE3) pLysS和Rosetta 2 (DE3)).
- 评估蛋白质溶解度和IMAC与Cu2+和Ni2+树脂的结合.
主要成果:
- CusF 聚变并没有改善 Efa PAP 或 mCherry 的可溶性.
- CusF-mCherry 主要积聚在不溶性分数中.
- 可溶性CusF-mCherry没有与Cu2+充电的IMAC树脂结合;与Ni2+充电的树脂结合是由于His标签.
结论:
- CusF并没有普遍提高蛋白质的溶解性.
- CusF可能无法连续地与基于铜的IMAC树脂结合.
- 测试多个可溶性标签对于优化重组蛋白质生产至关重要.
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