Generation of a vector suite for protein solubility screening

被引:18
作者
Correa, Agustin [1 ]
Ortega, Claudia [1 ]
Obal, Gonzalo [2 ]
Alzari, Pedro [3 ]
Vincentelli, Renaud [4 ]
Oppezzo, Pablo [1 ]
机构
[1] Inst Pasteur Montevideo, Recombinant Prot Unit, Montevideo 11400, Uruguay
[2] Inst Pasteur Montevideo, Prot Biophys Unit, Montevideo 11400, Uruguay
[3] Inst Pasteur, Unite Microbiol Struct, Paris, France
[4] Aix Marseille Univ, AFMB, CNRS, UMR7257, Marseille, France
来源
FRONTIERS IN MICROBIOLOGY | 2014年 / 5卷
关键词
recombinant proteins; solubility; expression; vector; cloning; high-throughput; SOLUBLE RECOMBINANT PROTEINS; CLOSTRIDIUM-THERMOCELLUM; FUSION PARTNER; EXPRESSION; PURIFICATION; CLONING; POLYPEPTIDES; TAG;
D O I
10.3389/fmicb.2014.00067
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
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.
引用
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页数:9
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