Preparation and Induction of Borrelia burgdorferi Cultures for Viral Particle Production

Begin with a culture of a genetically modified strain of Borrelia burgdorferi, a pathogenic bacterium.

This strain carries dormant bacteriophage DNA integrated into its genome, which has been engineered to include a target gene conferring antibiotic resistance.

Add the corresponding antibiotic and incubate the culture. The antibiotic resistance gene expresses a protein that inhibits the antibiotic, ensuring only bacteria harboring the gene proliferate.

Once the culture reaches the desired density, centrifuge the cells to pellet them and discard the supernatant.

Resuspend the pellet, introduce a chemical inducer, and incubate to induce DNA damage.

The resulting DNA repair response excises the bacteriophage DNA, initiating its replication, expression of structural proteins, and assembly of new virions carrying the antibiotic-resistance gene.

Centrifuge the culture again, discard the supernatant, and resuspend the cells in a fresh medium.

The bacterial cells containing mature virions are ready for further downstream applications.

Inoculate 150 microliters of the appropriate Borrelia burgdorferi clone into 15 milliliters of BSK in tightly-capped sterile conical centrifuge tubes for the transdu