Arjun and Patrick's Research Proposal

BoHV-4 Vaccine carrier

Use BoHV-4 (Bovine Herpes Virus 4) as a carrier to secrete a specific non-toxic antigen from a virus known to be harmful to humans, inject it into cell, and force the immune system to make antibodies against it.
Original Paper link: Establishment of a Bovine Herpesvirus 4 based vector expressing a secreted form of the Bovine Viral Diarrhoea Virus structural glycoprotein E2 for immunization purposes

Necessary components of expressed antigen:

Removal of the putative transmembrane domain
Addition of a 14 amino acids peptide at its carboxyl terminal
Immunoglobulin K signal peptide added to the amino terminal

Why are they needed?

Removal of putative transmembrane domain: Allows for the protein product to be secreted rather than inserted into membrane

Addition of 14 amino acids at carboxyl terminal: postulated and experimentally shown to increase secretion of protein; replaced transmembrane domain.

Immunoglobulin K signal peptide addition to amino terminal: Forces the immune system to recognize the secreted structure.

Why BoHV-4?

Non-pathogenic
Low probability of oncogenicity
Capable of carrying large amounts of foreign DNA
Ability to infect several different types of cells from different species
Ability to maintain transgene expression in either differentaited or undifferentiated cells

How to insert antigen into BoHV-4?

Recombination of modified antigen (as discussed before) DNA with BoHV-4 DNA, Cloned as bacterial artificial chromosome. Verify by using Kanamycin resistance gene in antigen cassette and selecting for it.

Predicted Outcomes

If all goes well, the use of BoHV-4 as a vehicle for the vaccine should allow for the cells to produce antibodies to the virus. If nothing goes according to plan, either the BoHV-4 is a bad vehicle or the antigen fails to allow the immune system to produce antibodies