Construction of an ASFV proteome library via multiple optimization strategies for high-throughput analysis

Area of Science:

• Virology
• Molecular Biology
• Biochemistry

Background:

• African swine fever virus (ASFV) is a complex DNA virus with over 160 encoded proteins.
• A comprehensive protein library is crucial for studying ASFV functions, antigenicity, and host interactions.

Purpose of the Study:

• To construct a high-coverage recombinant protein library for African swine fever virus (ASFV).
• To facilitate ASFV research, including protein function, antigenicity, vaccine development, and virus-host interactions.

Main Methods:

• Engineered glutathione S-transferase (GST)-tagged recombinant ASFV proteins expressed in a yeast host.
• Optimized codon usage, expression vectors, strains, and purification conditions to maximize protein yield.
• Developed a protein chip using the library to screen for ASFV-swine protein interactions.

Main Results:

• Achieved satisfactory protein yields for analytical applications, covering approximately 95% of the ASFV proteome.
• Successfully constructed and utilized a protein chip to identify interactions between ASFV and swine proteins (e.g., IRF3, p65, IκBα).

Conclusions:

• The ASFV protein library provides a foundational resource for understanding and controlling ASFV.
• The methods employed are applicable for generating other protein libraries for high-throughput biological applications.