[Preparation and identification of recombinant maxadilan]

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Engineering

Background:

• Maxadilan is a salivary protein with potential therapeutic applications.
• Efficient production of recombinant proteins is crucial for research and development.
• Gene engineering offers a pathway to synthesize complex biomolecules like Maxadilan.

Purpose of the Study:

• To produce recombinant Maxadilan using gene engineering techniques.
• To verify the identity and purity of the synthesized recombinant Maxadilan.
• To assess the biological activity of the recombinant Maxadilan.

Main Methods:

• Gene synthesis based on Maxadilan amino acid sequences.
• Construction of recombinant plasmid pKYB-MAX and transformation into Escherichia coli.
• Purification of MAX-intein-CBD fusion protein via chitin-affinity chromatography.
• Intein-mediated cleavage to release recombinant Maxadilan.
• Molecular weight determination using laser flight mass spectrometry.

Main Results:

• Successful synthesis and purification of recombinant Maxadilan.
• Confirmed molecular weight matching theoretical values.
• Demonstrated biological activity: significant enhancement of serum glucose concentration.

Conclusions:

• The gene engineering approach successfully produced functional recombinant Maxadilan.
• Recombinant Maxadilan exhibits biological activity, specifically increasing serum glucose levels.
• This method provides a viable strategy for producing Maxadilan for further study.