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Optimized gene synthesis and high expression of human interleukin-18
Ailian Li1, Zenichiro Kato, Hidenori Ohnishi
1Department of Pediatrics, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan.
Protein Expression and Purification
|December 19, 2003
Summary
Optimizing gene codons for human interleukin-18 (hIL-18) significantly boosted protein yield. This enhanced production method facilitates large-scale cytokine generation for structural and therapeutic applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- Human interleukin-18 (hIL-18) is a cytokine secreted by liver Kupffer cells and macrophages.
- Previous expression methods yielded low quantities of hIL-18.
- Skewed codon usage in native hIL-18 cDNA hinders efficient expression in Escherichia coli.
Purpose of the Study:
- To enhance the expression yield of human interleukin-18 (hIL-18).
- To address low protein yield issues caused by codon bias.
- To develop a scalable system for cytokine production.
Main Methods:
- Gene synthesis of mature hIL-18 with codons optimized for E. coli.
- Expression and purification of hIL-18 using the optimized gene sequence.
- Utilizing minimal medium for large-scale labeled protein production for NMR analysis.
Main Results:
- Optimized codon usage resulted in approximately a five-fold increase in hIL-18 protein yield compared to the native sequence.
- The system efficiently produces large quantities of labeled proteins suitable for NMR analysis.
- Demonstrated a simple and effective method for heterologous protein production.
Conclusions:
- Codon optimization is a critical strategy for improving the expression of cytokines like hIL-18 in E. coli.
- The developed production system offers a scalable and efficient approach for generating cytokines.
- This method has potential applications in producing other cytokines for structural and therapeutic research.