[논문]Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression

Chung, Jeong Min; Lee, Sangmin; Jung, Hyun Suk

doi:10.1016/j.pep.2016.06.010

Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression

Protein expression and purification, v.133, 2017년, pp.193 - 198

Chung, Jeong Min (Corresponding author.) , Lee, Sangmin , Jung, Hyun Suk

Abstract ▼ AI-Helper

Bacterial expression is commonly used to produce recombinant and truncated mutant eukaryotic proteins. However, heterologous protein expression may render synthesized proteins insoluble. The conventional method used to express a poorly soluble protein, which involves denaturation and refolding, is time-consuming and inefficient. There are several non-denaturing approaches that can increase the solubility of recombinant proteins that include using different bacterial cell strains, altering the time of induction, lowering the incubation temperature, and employing different detergents for purification. In this study, we compared several non-denaturing protocols to express and purify two insoluble 34 kDa actin-bundling protein mutants. The solubility of the mutant proteins was not affected by any of the approaches except for treatment with the detergent sarkosyl. These results indicate that sarkosyl can effectively improve the solubility of insoluble proteins during bacterial expression. (C) 2016 Published by Elsevier Inc.

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참고문헌 (29)

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Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression

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Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression

Abstract ▼ AI-Helper

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참고문헌 (29)

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