Multi-Compartment and Multi-Host Vector Suite for Recombinant Protein Expression and Purification.
Multi-Compartment and Multi-Host Vector Suite for Recombinant Protein Expression and Purification.
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DOI：
10.3389/fmicb.2018.01384
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发表时间：
2018
影响因子：
5.2
通讯作者：
Correa A
中科院分区：
文献类型：
--
作者：
Ortega C;Prieto D;Abreu C;Oppezzo P;Correa A
重组蛋白表达已成为基础和应用研究中的宝贵工具。在这一领域积累的知识使成千上万的蛋白质目标的表达在一个可溶的，纯的，和均匀的状态，必不可少的生化和结构分析。在过去的几十年中已经取得了很多进展，其中在评估不同的参数例如宿主、菌株和融合伴侣或启动子强度等之后，以可溶性方式表达具有挑战性的蛋白质。在这方面，我们以前已经开发了一个载体套件，允许不同的启动子和可溶性增强蛋白的评估，通过一个简单而有效的克隆策略。尽管如此，许多靶标的正确表达仍然难以捉摸，需要例如添加复杂的翻译后修饰和/或通过专门的区室。为了克服在使用单一亚细胞定位和单一宿主类型时发现的局限性，我们在此扩展了我们先前开发的载体套件以包括在不同细胞区室和细胞宿主中的重组蛋白表达的评价。此外，这些载体还允许评估用于提高靶蛋白产量的替代纯化策略。
Recombinant protein expression has become an invaluable tool in basic and applied research. The accumulated knowledge in this field allowed the expression of thousands of protein targets in a soluble, pure, and homogeneous state, essential for biochemical and structural analyses. A lot of progress has been achieved in the last decades, where challenging proteins were expressed in a soluble manner after evaluating different parameters such as host, strain, and fusion partner or promoter strength, among others. In this regard, we have previously developed a vector suite that allows the evaluation of different promoters and solubility enhancer-proteins, through an easy and efficient cloning strategy. Nonetheless, the proper expression of many targets remains elusive, requiring, for example, the addition of complex post-translation modifications and/or passage through specialized compartments. In order to overcome the limitations found when working with a single subcellular localization and a single host type, we herein expanded our previously developed vector suite to include the evaluation of recombinant protein expression in different cell compartments and cell hosts. In addition, these vectors also allow the assessment of alternative purification strategies for the improvement of target protein yields.
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影响因子：
6.4
作者：
Nozach H;Fruchart-Gaillard C;Fenaille F;Beau F;Ramos OH;Douzi B;Saez NJ;Moutiez M;Servent D;Gondry M;Thaï R;Cuniasse P;Vincentelli R;Dive V
通讯作者：
Dive V
影响因子：
6.4
作者：
Hatahet F;Nguyen VD;Salo KE;Ruddock LW
通讯作者：
Ruddock LW
DOI：
10.1007/978-1-4939-2205-5_2
发表时间：
2015-01-01
期刊：
INSOLUBLE PROTEINS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Correa, Agustin;Oppezzo, Pablo
通讯作者：
Oppezzo, Pablo
影响因子：
5.2
作者：
Correa, Agustin;Ortega, Claudia;Oppezzo, Pablo
通讯作者：
Oppezzo, Pablo
DOI：
10.1073/pnas.96.24.13703
发表时间：
1999-11-23
影响因子：
11.1
作者：
Bessette, PH;Åslund, F;Georgiou, G
通讯作者：
Georgiou, G