Expression of a human gene for interferon in yeast

Area of Science:

• Molecular Biology
• Biotechnology
• Recombinant Protein Production

Background:

• Human leukocyte interferon D (LeIF-D) is a crucial protein with therapeutic potential.
• Efficient and scalable production methods for LeIF-D are needed for its widespread application.
• Heterologous expression systems offer a promising avenue for producing complex proteins.

Purpose of the Study:

• To develop a system for the high-level expression of mature human leukocyte interferon D (LeIF-D) in yeast.
• To utilize the 5'-flanking sequences of the Saccharomyces cerevisiae alcohol dehydrogenase I gene for regulating LeIF-D production.
• To create a versatile expression plasmid functional in both yeast and Escherichia coli.

Main Methods:

• Cloning of the DNA sequence for mature LeIF-D into a yeast expression vector.
• Ligation of the LeIF-D coding sequence with the 5'-flanking sequences of the yeast alcohol dehydrogenase I gene.
• Transformation of yeast and Escherichia coli with the constructed plasmid for autonomous replication and selection.

Main Results:

• Successful construction of a plasmid enabling autonomous replication and selection in both yeast and E. coli.
• Transformed yeast cells synthesized significant quantities of biologically active LeIF-D.
• Expression levels reached up to 1 x 10(6) molecules of LeIF-D per yeast cell.

Conclusions:

• The developed expression system efficiently produces biologically active human leukocyte interferon D (LeIF-D) in yeast.
• This method provides a scalable platform for recombinant LeIF-D production.
• The findings support the use of yeast as a host for producing therapeutic proteins.