A novel method for increasing the expression level of recombinant proteins

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Expression

Background:

• Recombinant protein expression is crucial for gene function elucidation, target validation, and disease therapy development.
• Producing certain recombinant proteins can be challenging, hindering research and therapeutic advancements.

Purpose of the Study:

• To introduce a novel fusion protein expression system for overcoming challenges in producing difficult recombinant proteins.
• To demonstrate enhanced protein expression levels using a specific fusion partner.

Main Methods:

• A fusion partner, MTB32-C (carboxyl terminal fragment of Mycobacterium tuberculosis antigen MTB32), was fused to the N-termini of target genes.
• Recombinant protein expression was performed in Escherichia coli.
• mRNA levels were compared between fusion and non-fusion proteins.

Main Results:

• Significant enhancement of recombinant protein expression levels was achieved using the MTB32-C fusion partner.
• The fusion protein, including a 6xHis tag and MTB32-C, had an increased molecular weight.
• Enhanced expression may be regulated at translational or post-translational levels.

Conclusions:

• The MTB32-C fusion system is a powerful tool for enhancing recombinant protein expression, particularly for challenging proteins.
• Generated fusion proteins and derived antibodies have applications in antibody production, protein characterization, gene target validation, diagnostics, and therapeutics.
• The fusion partner can be removed, allowing for further study of protein structure, function, and drug screening.