AI Article Synopsis
- Heterologous protein expression in E. coli is widely used for generating recombinant proteins, but many proteins often end up as insoluble inclusion bodies.
- By analyzing 71 target proteins using different host strains and varying conditions (like purification tag location, promoter choice, and induction temperature), a successful protocol was developed.
- This protocol achieves a 77% success rate for producing soluble proteins, making it ideal for high-throughput screening aimed at structural studies.
Article Abstract
Heterologous protein expression in Escherichia coli is commonly used to obtain recombinant proteins for a variety of downstream applications. However, many proteins are not, or are only poorly, expressed in soluble form. High level expression often leads to the formation of inclusion bodies and an inactive product that needs to be refolded. By screening the solubility pattern for a set of 71 target proteins in different host-strains and varying parameters such as location of purification tag, promoter and induction temperature we propose a protocol with a success rate of 77% of clones returning a soluble protein. This protocol is particularly suitable for high-throughput screening with the goal to obtain soluble protein product for e.g. structure determination.
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| http://dx.doi.org/10.1016/j.pep.2011.08.030 | DOI Listing |
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