[Interleukin-2 gene cloning and expression in S. cerevisiae]

Area of Science:

• Molecular Biology
• Biochemistry
• Protein Expression

Background:

• Interleukin-2 (IL-2) is a crucial cytokine for immune response.
• Efficient and active production of recombinant IL-2 is essential for therapeutic and research applications.

Purpose of the Study:

• To develop a high-yield expression system for active human Interleukin-2 (IL-2) protein in yeast.
• To characterize the produced IL-2 protein, including its activity and potential post-translational modifications.

Main Methods:

• Polymerase Chain Reaction (PCR) amplification of the human IL-2 gene.
• Cloning into expression vectors (pUC12, pSK43SB, YEpHc8) using restriction enzymes and linkers.
• Recombinant protein expression in yeast transformants.
• Protein purification from yeast supernatant.
• Activity assays and sugar-specific staining for characterization.

Main Results:

• Successful amplification and cloning of the human IL-2 gene into an episomal expression vector.
• High-level expression and secretion of active human IL-2 protein by yeast.
• Purification of functional IL-2 protein with detectable carbohydrate components.
• Demonstration of IL-2 activity in the purified recombinant protein.

Conclusions:

• The developed yeast expression system provides a robust method for producing highly active, glycosylated human IL-2.
• This system facilitates efficient purification of recombinant IL-2 for potential therapeutic use.
• The presence of carbohydrate components suggests post-translational modification in yeast, relevant for protein function.