Design and application of expression constructs for FMDV serotype O structural proteins

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• Foot-and-Mouth Disease Virus (FMDV) poses a significant threat to livestock globally.
• Existing diagnostic and vaccine strategies require continuous improvement to address diverse and evolving FMDV strains.

Purpose of the Study:

• To design and validate flexible constructs for the recombinant expression of FMDV serotype O structural proteins.
• To enable efficient production of these proteins from currently circulating topotypes for enhanced control measures.

Main Methods:

• Degenerate primers were designed to target diverse FMDV topotypes for gene amplification (VP0, VP1, VP3).
• Overlap extension PCR was employed to integrate regulatory elements (T7 promoter, g10 leader, T7 terminator, RBS) for bacterial expression.
• Expression in E. coli was followed by SDS-PAGE and Western blotting for analysis.

Main Results:

• Successful amplification and cloning of VP0, VP1, and VP3 genes from FMDV serotype O.
• Recombinant proteins expressed at expected molecular weights (VP0: 34 kDa, VP1: 24 kDa, VP3: 22 kDa).
• High protein yield and purity confirmed by SDS-PAGE and Western blotting.

Conclusions:

• A robust platform was established for the efficient recombinant expression of FMDV structural proteins.
• The developed constructs and methods are adaptable for diagnostic and vaccine development.
• This work provides a valuable tool for improving FMDV control strategies through better diagnosis and vaccination.