Effect of osmotic stress and heat shock in recombinant protein overexpression and crystallization |
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Authors: | Oganesyan Natalia Ankoudinova Irina Kim Sung-Hou Kim Rosalind |
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Institution: | Bio-Rad Laboratories, Life Science Group, 2000 Alfred Nobel Drive, Hercules, CA 94547, USA. |
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Abstract: | Overexpressed recombinant proteins in bacteria often tend to misfold and accumulate as soluble aggregates and/or inclusion bodies. A strategy for improving the level of expression of recombinant proteins in a soluble native form is to increase the cellular concentration of osmolytes or of chaperones. This can be accomplished by growing the bacterial cells in the presence of high salt, sorbitol, and betaine as well as exposing the cells to a heat shock step. Our results suggest that by growing the cells under varied conditions one may be able to express targets as soluble proteins (from previously insoluble targets) and to improve the chances of their crystallization. |
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Keywords: | Recombinant protein Solubility Crystallization Osmotic stress |
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