重组DNA
溶解度
增强子
融合蛋白
计算生物学
克隆(编程)
一套
载体(分子生物学)
表达式向量
生物
计算机科学
化学
生物化学
基因表达
基因
考古
有机化学
历史
程序设计语言
作者
Agustín Correa,Claudia Ortega,Gonzalo Obal,Pedro M. Alzari,Renaud Vincentelli,Pablo Oppezzo
标识
DOI:10.3389/fmicb.2014.00067
摘要
Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions.
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