[Construction and Expression of RNase-Resisting His-Tagged Virus-Like Particles Containing FluA/B mRNA]

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• Accurate detection of Influenza A and B viruses is crucial for public health.
• Existing RNA detection methods require reliable standards and controls.
• Virus-like particles (VLPs) offer a promising platform for molecular diagnostics.

Purpose of the Study:

• To engineer VLPs encapsulating Influenza A/B mRNA.
• To establish these VLPs as a stable RNA standard and control for influenza detection.
• To optimize VLP production and purification for diagnostic applications.

Main Methods:

• Genes for MS2 bacteriophage coat protein and maturase were cloned into a D-pET32a vector.
• A 6-histidine tag was introduced to the MS2 coat protein.
• Influenza A and B gene fragments were inserted downstream, creating a recombinant plasmid.
• Recombinant plasmid was expressed in BL21 E. coli, and His-tagged VLPs were purified via Ni+ chromatography.

Main Results:

• Purified VLPs successfully encapsulated Influenza A/B mRNA.
• VLPs were detectable by RT-PCR but not standard PCR, confirming RNA encapsulation.
• VLPs demonstrated stability for at least 3 months at 4°C and -20°C.
• His-tagged VLPs exhibited enhanced stability and simplified purification.

Conclusions:

• Engineered His-tagged MS2 VLPs effectively encapsulate Influenza A/B mRNA.
• These VLPs serve as stable and reliable RNA standards and controls for influenza detection.
• The developed VLP system provides a valuable tool for improving the accuracy and consistency of influenza diagnostics.