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Updated: Jun 28, 2025

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

对基于光蛋白的方法进行新的研究,以确定E. coli重组蛋白表达的最佳编码序列

Artur I Zabolotskii¹, Natalia S Riabkova¹

¹Lomonosov Moscow State University, Moscow, Russia.

Biotechnology journal

|April 16, 2024

概括

优化基因中的同名编码子.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
遗传学是一个遗传学.

背景情况:

遗传密码的退化允许一个氨基酸有多个编码子.
同义代码子选择影响蛋白质表达,特别是在大肠杆菌中.
在5'-end的mRNA二次结构可以阻碍翻译启动.

研究的目的:

开发一种高效的方法来优化蛋白质表达,使用同义代码子替代.
为了克服in silico基因优化的局限性.
增强细菌中表达不良的蛋白质的表达.

主要方法:

创建一个部分随机的图书馆的表达式构造与选定的同义变体.
感兴趣的基因与光蛋白记者基因的融合.
基于记者信号强度选变异,以识别高表达性候选者.

主要成果:

证明了三种蛋白质的 prokaryotic 表达的显著增加:犬类的 cystatin C,人类的 BCL2 相关的 athanogene 3 和人类的心脏 I.
验证了选方法的有效性,以识别最佳的同名密码体变异.
通过这种方法实现了蛋白质产量的大幅改善.

结论:

为优化细菌中的蛋白质表达而开发了一种新,简单和具有成本效益的方法.
这种方法为表达不良的蛋白质提供了一个有效的替代品,而不是在中优化.
随机图书馆的选为基因工程和蛋白质生产提供了一个强大的工具.

关键词:

美国大肠杆菌 (E. coli).表达式优化表达式优化在Silico优化中的优化合成图书馆合成图书馆

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