The use of outer membrane proteins as an exposure surface for foreign antigens in AIDS vaccine methodology and AIDS diagnostics

Abstract

A live recombinant bacteria or virus with HIV determinants exposed at the outermembrane is one strategy for AIDS vaccine development. Two HIV determinants that showed neutralization capacity in in- vitro experiments were tested for their expression in PhoE outer membrane protein of E coli K12. Determinants, located on HIV envelope glycoprotein gp120, were inserted into the unique Nru I restriction site created in the fourth exposed region of PhoE. Partial insertion of one of the envelope determinants into the PhoE gene, having an inserted length of 27 amino acid residues, was succesfully done. The level of expression of the hybrid protein was however decreased compared to literature. Correct insertion of the other HIV derterminant, having an inserted length of 22 amino acid residues, couldn't be well determined yet. Experiments to enhance protein synthesis with lamoda promotor controlled expression were not succesful yet. Further research has to be done to improve expression of HIV determinants in PhoE as well as other outer membrane proteins, like Lam B, have to be tested as expression vector.<br>