Bacterial expression and purification of biologically active mouse c-Fos proteins by selective codon optimization

Area of Science:

• Molecular Biology
• Protein Expression
• Biochemistry

Background:

• Mouse c-Fos protein is a key transcription factor involved in cellular processes.
• Achieving high-level expression of eukaryotic proteins in prokaryotic systems like E. coli can be challenging.
• Codon optimization is a common strategy to improve protein expression.

Purpose of the Study:

• To develop a simple strategy for high-level expression of mouse c-Fos protein in E. coli.
• To investigate the impact of selective codon optimization on protein yield.
• To confirm the biological activity of the expressed and purified mouse c-Fos protein.

Main Methods:

• Selective codon optimization of ten arginine codons in the basic region of mouse c-Fos.
• Expression of the optimized mouse c-Fos gene in E. coli.
• Purification of the expressed c-Fos protein.
• Assay of biological activities to confirm protein function.

Main Results:

• Successfully achieved high-level expression of mouse c-Fos protein in E. coli.
• Purified c-Fos protein to near homogeneity.
• Demonstrated the biological activity of the purified mouse c-Fos protein through functional assays.

Conclusions:

• Selective codon optimization is an effective strategy for enhancing mouse c-Fos protein expression in E. coli.
• The expressed and purified mouse c-Fos protein retains its biological activity.
• This method provides a viable approach for producing functional mouse c-Fos protein for research purposes.