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Updated: Jun 24, 2025

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一个系统的分析的亲和度标签在haloarchaeal表达系统,Haloferax volcanii用于蛋白质净化

Ram Karan^1,2, Dominik Renn², Thorsten Allers³

¹Department of Microbiology, University of Delhi, South Campus, New Delhi, India.

Frontiers in microbiology

|June 14, 2024

概括

在Haloferax volcanii (H. volcanii) 中优化净化标签可以增强极端性蛋白质的产生. N-终端8xHis-tag或Strep-tag®II显著提高了蛋白质的产量和纯度. 在这个友系统中.

科学领域:

生物技术是生物技术.
蛋白质表达方式
极端爱好研究极端爱好研究

背景情况:

极端性蛋白质提供了独特的优势,但很难在像大肠杆菌这样的标准宿主中表达.
错误折叠和聚合是异质蛋白表达的常见挑战.
火山 (Haloferax volcanii) 是一个有前途的性宿主,可以表达具有挑战性的蛋白质.

研究的目的:

研究不同净化标签 (可切割/不可切割,N-/C-终端) 对H. volcanii.蛋白质生产的影响.
确定最佳的标记策略,以提高蛋白质产量和纯度.
为了评估各种记者和功能蛋白的标签性能.

主要方法:

sfGFP,mCherry和ADH与各种净化标签 (8xHis,Strep-tag®II,Twin-Strep-tag®) 的融合.
在性宿主H. volcanii.中标记蛋白的表达.
使用生物化学分析分析蛋白质的生产,纯度和产量.

主要成果:

N-终端8xHis-tag或Strep-tag®II显著改善了sfGFP的生产,纯度和产量.
最佳的标签放置因蛋白质而异:mCherry的C端,ADH的N端.
在N端的8xHis-tag和Twin-Strep-tag®的同时表达进一步提高了所有测试蛋白质的产量.

结论:

净化标签的战略选择和放置对于H. volcanii.中的蛋白质表达成功至关重要.
N-终端标签,特别是8xHis和Strep-tag®II,在提高该系统中的蛋白质生产方面非常有效.
这些发现为优化生物技术应用中性宿主中的重组蛋白质生产提供了一个框架.

关键词:

它们是Haloferax volcanii (火山).他的标签是他的标签.链杆菌标签®IIII 的标签.考古学是指古物质 (archaea) 是指古物质.生物工程是生物工程.他们是极端爱好者 (extremophiles).这种类型的类动物是halofil.净化标签是指净化标签.

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