Using Maltose-Binding Protein to Improve Protein Expression in E. coli
Author Information
Author(s): Hewitt Stephen N., Choi Ryan, Kelley Angela, Crowther Gregory J., Napuli Alberto J., Van Voorhis Wesley C.
Primary Institution: Seattle Structural Genomics Center for Infectious Disease, University of Washington
Hypothesis
Can maltose-binding protein (MBP) fusion improve the expression and solubility of previously non-expressing proteins in E. coli?
Conclusion
The study found that MBP can significantly enhance the soluble expression of proteins that previously could not be expressed.
Supporting Evidence
- 72% of the clones showed detectable expression after using the MBP fusion strategy.
- 62% of the expressed proteins were soluble.
- Further refinements in MBP tagging may improve the use of MBP-fusion proteins in crystallographic studies.
Takeaway
Scientists tried to help bacteria make proteins by sticking a helper protein called MBP to them, and it worked for many of the proteins that wouldn't normally show up.
Methodology
The study involved cloning genes into an MBP-fusion vector and analyzing expression and solubility through high-throughput nickel-affinity binding.
Limitations
The solubility of most proteins was marginal to poor after cleavage of the MBP tag.
Digital Object Identifier (DOI)
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